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Cleaning the Agilent 3100 OFFGEL Fractionator

Contents

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7. B C HIV I V 128 OFFGEL Agilent 3100 OFFGEL Fractionator M Benutzerhandbuch ee e O o o 270 Sicherheit x Sicherheitshinweise 130 Dieses Kapitel enth lt wichtige Sicherheitshinweise zu dem Agilent 3100 OFFGEL Fractionator RE Agilent Technologies 129 Sicherheit Sicherheitshinweise Die folgenden allgemeinen Sicherheitsma nahmen m ssen in allen Phasen des Betriebs der Wartung und der Reparatur des Agilent 3100 OFFGEL Fractionator Die Bediener sollten vor der Installation dem Betrieb und der Wartung des Gerats alle Sicherheitsanweisungen sorgfaltig durchgelesen und verstanden haben Die Nichtbeachtung dieser Vorsichtsma nahmen oder spezifischer Warnungen die an anderen Stellen in diesem Handbuch enthalten sind verst t gegen die Sicherheitsstandards f r Konstruktion
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10. Revision Build Revision Build Figure 38 Current Firmware Information screen 2 Above screen shows the current firmware information 3 Select the Revision History tab This window shows details of the used main board and the update history in the module see Figure 37 84 OFFGEL User Manual OFFGEL User Manual Maintenance 6 Firmware Update Firmware Update Tool File Action Options Help QE Ses All Instruments E 3100 OFFGEL 1 Current Firmware Update InformSNgg_ Revision History G31004 DE 00000070 AGILENT TECHNOLOGIES PrimaryLoader 6 1 I 4 6 2010 10 17 46 AM B 06 30 002 4 6 2010 10 09 27 AM B 01 08 001 Figure 37 Revision History Information screen 4 Select the Update Information tab In this window you can select the firmware for the main and resident part in the module If no firmware is listed when you click on the file selection button correct the firmware folder locations see Figure 30 on page 81 5 Activate the check boxes for the firmware This firmware part is added to the update list 85 6 Maintenance Firmware Update Firmware Update Tool sf 3100 OFFGEL 1 Current Firmwal Update Information ReBision History Main System Qe Filename Revision Build 31004 U630 003 dlb af G31004 DE00000070 Resident System Update Filename Revision Build r G31004 DE00000070 Main 3100 OFFGEL 1 Figure 38 Update Information screen
11. This product complies with the WEEE Directive 2002 96 EC marking requirements The affixed label indicates that you must not discard this electrical electronic product in domestic household waste Product Category With reference to the equipment types in the WEEE Directive Annex I this product is classed as a Monitoring and Control EIE instrumentation product Do not dispose of in domestic household waste To return unwanted products contact your local Agilent office or see www agilent com for more information Sound Emission This statement is provided to comply with the requirements of the German Sound Emission Directive of January 18 1991 OFFGEL User Manual 23 Site requirements and specifications This product has a sound pressure emission at the operator position lt 70 dB A Sound Pressure Lp lt 70 dB A At Operator Position Normal Operation According to ISO 7779 1988 EN 27779 1991 Type Test Instruction for Use The Agilent 3100 OFFGEL Fractionator is designed and tested as general purpose electrical equipment for laboratory use It is designed and tested for research use only and not for use in diagnostic procedures CAUTION If the equipment is used in a manner not specified in this manual the protection provided by the equipment may be impaired 24 OFFGEL User Manual Site requirements and specifications 2 Physical Specifications of the Agilent 3100 OFFGEL Fractio
12. 6 Select the Start Update button All selected modules are updated The update status is shown at the bottom of the screen In case you also update the resident Firmware this will be done first followed by the main firmware NOTE When the main firmware is being updated the Status LED of the instrument is red and us blinking 3100 0FFGEL 1 Updating firmware please wait LE 00 00 52 Figure 39 Update is running 86 OFFGEL User Manual Maintenance 6 If you cancel an update the module remains in the current mode resident or main depending when the process was stopped It cannot be used until the update process has been restarted At the end of the update a report is shown The update status is either PASS successful or FAILED not successful see Figure 40 If the update failed the Protocol file contains more information see The Log file on page 87 Firmware Update Tool Update Summary The update process has been logged to the following file C Program Files gilent LC Firmware Update Tool FwReport tat Module System Result G31004 DE00000070 Main SUCCEEDED Figure 40 Report Screen 7 If desired open and print the Protocol file 8 Press OK to continue 9 Perform a power cycle of the instrument The Log file This file is the verification that the firmware update was completed successfully or in the case of problems the log file may give some specific information Press
13. Do not disturb the firmware process Any interruption will lead to incomplete firmware programming Instruments in such status are not functional OFFGEL User Manual 89 6 Maintenance 9 Populate the web site with the instrument user interface in your browser again Confirm the changed main firmware in the About screen 10 Remove the USB stick from instrument To perform an update of the Resident part of the Firmware create a folder X OFFGEL RESIUP on the USB stick provide the firmware file and follow steps 6 11 An example for a typical Resident Firmware file name is res_B623_001 dlb There is no way to check the resident firmware from the web interface Please use the Firmware Update Tool to confirm the success Main firmware is automatically downgraded when the USB stick is mounted on startup and the directory X OFFGEL UPDATE does not exist or is empty and exactly one dlb file for the Agilent 3100 OFFGEL Fractionator with a firmware version older than the current version is present in the directory X OFFGEL DOWNDATE Resident firmware is automatically downgraded when the USB stick is mounted on startup and the directory X OFFGEL RESIUP does not exist or is empty and exactly one dlb file containing a resident firmware version older than the current version is present in the directory X OFFGEL RESIDOWN If the USB stick contains both a folder for resident and a main system the device will first update or downgrade
14. Edit Stop Figure 43 Typical current pattern on tray when every second lane is filled with water No currents should flow in the empty tray lanes lt 1 0 uA If this is the case proceed with step 10 Otherwise stop here because you may have contaminated electrodes electrode connectors or trays Clean and carefully dry the electrodes trays and the electrode connectors as described in Cleaning the Agilent 3100 OFFGEL Fractionator on page 70 and start this test again at step 6 Typical values for currents are between 20 uA and 50 uA and one lane has to be at maximum current of 50 uA representing the current limit if the voltage is just at 800 V or below 800 V In case the voltage is constantly at 800 V representing the voltage limit a current lower than 50 uA might be present in all lanes Differences in current from lane to lane are natural and might be due to variation in the resistance of the electrode surface The voltage has to be greater than 500 V The absolute value for the voltage is dependent on the conductivity of the deionized water used and is not a criterion for the proper function of the instrument The more salt ions the water contains the lower is the voltage value 10 Open the top cover and pipette 4 ml deionized water 18 MOhm m into lanes B D F and H on each tray 11 Close the top cover OFFGEL User Manual Troubleshooting 7 Troubleshooting the Instrument 12 You should now see the a typical cu
15. Example of log file with successful update If you encounter any problems with the firmware update feel free to send an e mail with your findings to our technical support specialists http www chem agilent com scripts cag techsupport asp or contact your local Agilent Technologies representative Attach your log file and details of your system revision and errors Firmware Change via USB Stick A convenient alternative to the Firmware Update Tool is the instrument Firmware update via USB stick Prerequisite is the Main Firmware on the instrument is revision B 06 30 or higher Please check in the About screen of the web interface of your instrument see Figure 24 on page 64 if this is installed The USB memory stick used must be FAT 16 formatted and without encryption maximum size is 2 GB See List of Spare Parts on page 156 In general the suitability of an USB stick can be check by inserting it simply to OFFGEL User Manual Maintenance 6 the port at the rear of instrument Upon recognition the browser interface will show the USB stick symbol in the status screen In case the USB memory stick is FAT 32 formatted the instrument will attempt to format it to FAT 16 The file browser of your computer operating system also allows you to format USB sticks FAT 16 is typically noted simply as FAT Steps for Firmware Change via USB 1 Download the Agilent 3100 OFFGEL Fractionator firmware from the Agilent web site http ww
16. pl obtained acts as an additional validation parameter No MS incompatible additives used in fractionation buffer The instrument can also be operated in the traditional in gel mode IEF Fractionation of up to 16 samples in parallel on 2 trays accommodating 8 samples each Broad loading capacity range from 50 ug sample load up to 5 mg sample load Two independent power supplies allow two different methods to be run at the same time Easy method setup and monitoring of important fractionation parameters and settings such as voltages and currents during a run Online current control for each individual sample or gel strip during fractionation Viewing of up to three run parameters in an online plot Easy export of important run parameters of a fractionation via LAN to the control PC or to a USB stick for further analysis Automated storage of up to 1200 data files including important run parameters on the compact flash card Monitoring of all operations and error events using the self updating logbooks OFFGEL User Manual Product overview 1 Instrument and Accessories Instrument and Accessories Figure 2 gives an overview of the complete Agilent 3100 OFFGEL Fractionator system Top Cover Cooling Platform Serial number Figure 2 Instrument overview OFFGEL User Manual 17 1 Product overview Anode Connector Cathode Connectors Trays Cooling Platform 18 The instrument has two individual h
17. o SS 16 L 1 2 3 4 5 6 7 8 3 10 Figure 46 Beta lactoglobulin Protein Test Sample in well 3 of a 12 cm pH 3 10 IPG strip Gel like image from an Agilent 2100 bioanalyzer analysis using the Protein 80 kit of wells 1 9 Example 2 Current Drop During Fractionation Figure 47 shows an example where the current see arrow dropped close to zero after approximately 30 minutes of the run In addition the volt and power signal is highly fluctuating OFFGEL User Manual 107 7 7 108 Troubleshooting Exp 52 nativ buffer 6000 250 kVHours Volt Power 200 n m i NL N mM CIMA i e AI 5000 150 3000 V uAh uA mW kV h time h Figure 47 Example 2 current drop during fractionation The root cause of this focusing failure was that the IPG strip was dried out at one position so that the conductivity was lost Example 3 Reduced Electrical Charge in One Lane Figure 48 shows an example of a fractionation of eight samples in parallel and just highlights the different electrical charges in uAh that were measured throughout the focusing for each lane The signal of electrical charge for lane F see arrow is clearly lower than for the other lanes Indeed it could be verified in a follow up experiment that the focusing of lane F was impaired OFFGEL User Manual Troubleshooting 7 Troubl
18. Fig 43 allows to setup and to review LAN configuration of the instrument The active IP address that is currently in use is highlighted by a blue corona On this tab you can also trigger module identification 1 User LAN Status Method Plot Logbook Results About P Configuration idie mm 2 Default LAN configuration 3 Set Identify Lan Configuration user settable address oS e Set identify Lan Configuration default address IP Address 192 168 Subnet Mask 255 255 Default Gateway 255 255 LAN Configuration Operation 5 The Agilent 3100 OFFGEL Fractionator has two LAN configurations stored They can be used alternatively The selection which of the stored LAN configurations is used upon the next instrument start is done by configuration dip switches at the rear of the instrument Any change in the LAN configuration is applied only after the instrument has been restarted Configuration dip switches 7 and 8 up while all other switches are down configures the instrument to start with the default IP address 192 168 254 11 Only configuration dip switches 7 up while all other switches are down configures the instrument to start with the user settable static IP address A User LAN Configuration with a settable address is visible in the upper panel Figure10 The user defined address must be a static IP address DHCP Dynamic Host Configuration Protocol is not supported The user sett
19. 3 Press the Start button to start the fractionation You will be able to monitor the voltage applied and the current flowing through the individual lanes from the status page It is recommended to monitor the startup phase in order to correct for example for missing connectivity You can protect the instrument from an unwanted access of a third user by the lock function see Instrument lock on page 45 A blue light below the anode connector of the respective tray indicates that the instrument is running A blinking blue light indicates that the fractionation is finished Both trays can be used independently If the top cover is opened the fractionation pauses until the top cover is closed again 4 Upon finished fractionation you can stop the run and retrieve the fractions 68 OFFGEL User Manual Agilent 3100 OFFGEL Fractionator M User Manual 000 ee 6 e Maintenance Cleaning the Agilent 3100 OFFGEL Fractionator 70 Firmware Update 76 This chapter describes cleaning procedures that should be used to do maintenance on your instrument at regular intervals or before every use In addition it provides instructions on how to update the firmware Only perform maintenance on the Agilent 3100 OFFGEL Fractionator as specified in this chapter Other maintenance and repairs must be done by Agilent Technologies trained personnel Unauthorized maintenance can be dangerous and damage is not covered by warranty RE Agilent Technologi
20. 92 T temperature 42 temperature fluctuations 22 top cover 17 75 Index tray 19 66 71 cleaning 71 lock 45 preparing 66 set 156 troubleshooting application 102 instrument 92 U un install firmware update tool 77 unpacking 32 update info screen 86 USB connection 34 USB stick 57 156 firmware update 88 folder directories 57 user interface 40 V vapores inflamables 149 voltage selector 22 W warning 114 120 126 136 142 weight 25 158 www agilent com In This Book Technical information about the Agilent 3100 OFFGEL Fractionator is given Site and equipment requirements Fractionation principles Installation and configuration Using the browser interface Maintenance and Troubleshooting Spare parts and consumables Safety and related information Agilent Technologies 2010 Printed in Germany 04 2010 0001 Agilent Technologies
21. EAU LIL of God T Nr Agilent 3100 OFFGEL ui Ex 0 UT Safety Class I E 120 OFFGEL Agilent 3100 OFFGEL
22. Modify Instrument Support G1323B active not active Help Help About revision of program Table 7 Functions of Buttons Button Description Connect to module system Disconnect from module system Select for update of all modules of a system Start the update for all selected modules MER M Add instrument to the list Delete instrument from the list T Modify instrument name or the connection can be changed Configure directories of the firmware files and report Open protocol file logbook with details of the actions made during the update e a amp Configure the Firmware Update Tool 1 Select the button Configure Directories and browse for an alternative location of the firmware files e g E Fwupgrad LC You can move a firmware file OFFGEL User Manual Maintenance Firmware Update as well to the default location C Program Files Agilent LC Firmware Update Tool ES Firmware Update Tool File Action Options Help e 2 v gt x SEE OE ipa Figure 29 Select the button Configure Directories Configure Directories x Set the directory that contains the firmware files and specify the location of the protocol file Firmware directory E FwupgracLC Protocol file C Program Files gilent LC Firmware Update Tool FWReport txt TN OK Cancel Figure 30 Configure Directories The Protocol file is a text file that gives the complete status o
23. USB memory stick which is incompatible due to the formatting it will try to format memory to FAT 16 Please be aware that any information present on the stick will be lost 56 OFFGEL User Manual Operation 5 Reviewing Result Files Result files are either transferred to your control PC by direct download using the link from the result tab and the browser function Save Target as or they were transferred to the USB stick mounted to the 3100 OFFGEL fractionators In order to review them it is reasonable to store them in a typical location of the Computer used for reviewing them such as under MyDocuments How to find the exported files on the USB stick When exporting results or transferring methods to an USB stick the following directories are generated Folders Size Type Date Modified KINGSTON F 1KB Microsoft Office Excel 8 16 2006 17 57 E OFFGEL O METH E Gres E C3 00000041 E 2006 E 06 a 23 6 13 so B 14 os 4 3 28 E 14 Qi Figure 18 Folder directories on the USB stick The METH folder contains all exported methods The RES folder contains all exported results The subdirectory of the RES folder starts with an instrument folder in the name of the instrument serial number The following directories follow the structure of the export menu year month day hour minute Figure 17 on page 54 The csv file in the minute folder finally contains all run parameters
24. all operatore e proteggere gli strumenti da eventuali danni Sicurezza chimica e biologica Lavorando con i solventi osservare sempre le norme di sicurezza in vigore indossare gli occhiali e gli abiti antinforturnistici come descritto nella scheda tecnica del materiale fornita dal rivenditore specialmente se si utilizzano solventi tossici o pericolosi ATTENZIONE 144 Manuale per l utente del frazionatore OFFGEL ATTENZIONE Sicurezza Se si intende utilizzare campioni tossici o radioattivi sullo strumento responsabilita dell utente garantire che tutte le norme di sicurezza in vigore linee guida precauzioni e pratiche vengano rispettate adeguatamente Questo anche in caso di manipolazione di organismi geneticamente modificati Informarsi presso il responsabile della sicurezza del laboratorio sul livello di protezione richiesta per l applicazione in uso e sui procedimenti di decontaminazione e sterilizzazione da seguire in caso i fluidi fuoriescano dai contenitori Osservare attentamente tutte le avvertenze riportate sui contenitori delle soluzioni originali prima dell uso Poich perdite fuoriuscite e versamenti di campione possono produrre aerosol seguire procedure di sicurezza adeguate I coperchi del sistema Frazionatore OFFGEL Agilent 3100 non sono stati progettati per fungere da tenute biologiche per il contenimento di liquidi o aerosol Maneggiare i fluidi corporei con estrema cura poich possono trasmettere
25. des phases d utilisation d entretien et de r paration du Fractionneur Agilent 3100 OFFGEL Toutes les consignes de s curit doivent tre lues et assimil es avant l installa tion l utilisation et toute op ration de maintenance de l instrument La non observation de ces consignes ou des avertissements sp cifiques indiqu s dans ce manuel constitue une violation des normes de s curit relatives la concep tion la fabrication et l utilisation normale de l instrument Agilent Technologies ne saurait tre tenu pour responsable en cas de non observation de ces consignes par le client Cet avertissement vous signale une situation susceptible d entrainer des blessures ou une d t rioration de l quipement N allez pas plus loin tant que vous n avez pas pleinement compris et rempli les conditions indiqu es ATTENTION Cet avertissement vous signale des situations pouvant mettre en p ril les fonctionnalit s ou les performances de l appareil Fractionneur Agilent 3100 OFFGEL ou conduire des pertes de donn es N allez pas plus loin tant que vous n avez pas pleinement rempli les conditions indiqu es S curit de l instrument Il s agit d un instrument de classe quip d une borne de mise la terre de protection qui a t fabriqu et test selon les normes internationales de s curit 136 Manuel de l utilisateur OFFGEL S curit Cordons secteur Diff rents cordons secteur sont propos s en
26. describes an example of the tray preparation for one IPG strip 1 Place the IPG strip in a new tray lane with the gel side up The low pH side Anode marked with a must be at the left where the fixed electrode will be attached 2 Take two electrode pads with tweezers wet them with rehydration buffer and place one at each end of the IPG strip so that it connects to the swelled gel 3 Attach the electrodes to the tray exactly as described in the OFFGEL Kit Manual 5969 1582 If necessary reposition the IPG strip with the electrode pads so that the fixed electrode connects to the left electrode pad 4 Place the tray into the instrument 5 Cover the complete tray lane with mineral oil using 1 5 ml mineral oil per IPG strip 66 OFFGEL User Manual Operation 5 Performing a Fractionation NOTE Distribute the mineral oil evenly across the whole tray lane by pipetting droplets across the lane Make sure that no mineral oil is spilled out of the tray 6 Start the run as described in Starting a Run on page 68 OFFGEL User Manual 67 5 Operation Starting a Run 1 To start the fractionation make sure you have configured the cooling platform according to your needs For details refer to General Functions on page 42 2 From the Status screen of the connected instrument select the Method in the drop down menu of the tray you will use The run parameters of the loaded method are visible on status screen
27. di alimentazione forniti da Agilent Technologies Funzionamento Prima di alimentare il sistema seguire attentamente le istruzioni per l installazione affinch lo strumento possa funzionare in sicurezza Non togliere mai i coperchi della strumentazione durante il funzionamento Prima dell accensione tutti i terminali protettivi le prolunghe gli autotrasfor matori e qualsiasi dispositivo collegato devono essere collegati ad una presa dotata di messa a terra Qualsiasi interruzione della messa a terra protettiva e una potenziale causa di scosse elettriche in grado di provocare lesioni gravi alle persone Quando possibile che la protezione sia danneggiata lo strumento deve essere scollegato e reso inutilizzabile Non effettuare operazioni di manutenzione senza aver staccato la corrente Non sostituire i componenti dello strumento Non utilizzare lo strumento in presenza di gas o vapori infiammabili L uso di qualsiasi tipo di apparecchiatura elettrica in ambienti di questo tipo pu comprometterne la sicurezza Non installare parti di ricambio o effettuare modifiche non autorizzate allo strumento Manuale per l utente del frazionatore OFFGEL 143 Sicurezza Simboli di sicurezza Tabella 13 Simboli di sicurezza utilizzati per gli strumenti e per la documentazione tecnica Simbolo Descrizione L apparecchiatura e contrassegnata da questo simbolo quando l utente deve consultare il Manuale per l utente per evitare danni
28. in case of serious issues LJ Posi Status Method Plot Logbook Results Config Agilent Technologies 3100 Offgel Fractionator Firmware version U 06 30 0009 Visit the 3100 OFFGEL fractionator web page Licences MIT License jQuery JavaScript Library jQuery UI jQuery JSON Plugin jQuery cookies 2 2 0 js jQuery flot js Apache License 2 0 excanvas js Codeplex jQuery Form Validators http validator codeplex com license For service purposes download DiagBuffers txt Download Agilent 1100 1200 Series Firmware Updste Tool Figure 24 About the Agilent 3100 OFFGEL Fractionator Operation 5 Performing a Fractionation OFFGEL User Manual Fractionations using the Agilent 3100 OFFGEL Fractionator can be performed in the OFFGEL mode or in the standard IPG IEF mode in gel mode The table lists the default methods delivered with the system and ready to use with a cooling platform temperature of 20 C Table 6 Default methods Method name recommended use 0G24PR01 0G24PE01 0G12PR01 0G12PE01 IG 24 00 IG 13 00 DIAGNOSE OFFGEL default method for proteins for use with 3100 OFFGEL High Res Kit pH 3 10 24 well frames OR for use with 3100 OFFGEL High Res Kit pH 4 7 24 well frame OFFGEL default method for peptides for use with 3100 OFFGEL High Res Kit pH 3 10 24 well frames OR for use with 3100 OFFGEL High Res Kit pH 4 7 24 well frame OFFGEL default method for proteins for use
29. mechanism from the browser Internet Explorer is not available on the iPod A different way to access and analyze the result files is to copy them to a connected USB stick To use this function select a day month or year by clicking with the mouse to a day within the desired period Click for example on Day at the USB Export selection This will export or copy all files of the selected day to the USB stick Pressing Month copies all of the currently selected month and Year all files of the selected year The progress of copying is shown in the USB progress bar The compact flash card which is inserted at the rear of the instrument has a limited capacity It can store results from a period up to approximately 3 years The storage capacity is dependent on the file size and is thus method dependent When the compact flash card is full the oldest data file is overwritten automatically Therefore old files might become unavailable It is recommended to archive result files on a different location Since USB memory sticks may vary from vendor to vendor or from type to type incompatibilities can occur In general USB memory sticks from Sandisk and Kingston were used with success The USB memory sticks must be FAT 16 formatted and without encryption maximum size is 2 GB See Firmware Change via USB Stick on page 88 A compatible USB stick is included with the shipment OFFGEL User Manual 55 5 Operation In case the instrument detects an
30. method name to store the setpoints to Functions in the timetable pane are Set Takes over the set values for the currently selected point in the timetable graph Add Uses the set values to create a new point in the timetable Delete Removes the currently selected point from the time table Factory installed default methods cannot be changed or deleted Please save changed method under a different name Default methods on the internal disk are part of the system and cannot be deleted A method that is currently loaded to one of the trays or is being executed cannot be deleted from the method list OFFGEL User Manual Operation 5 Displaying Data Graphically The plot tab Figure 15 allows viewing up to 4 signals which can be Volt Volt Hours Voltage Limit Current Limit and Current per lane The signals are available from the Signal Selection Menu The scaling source of the Y Axis is selectable by clicking onto the data series in the Graph area or reselecting it from the menu The view area can be zoomed by clicking to the graph area of interest and using the mouse wheel Clicking into the graph area and dragging allows moving the zoomed area to another section Unzoom is available under Display Options A stacked and overlay view onto the signal is possible as well as a switch between Tray I and Tray II Status Method Plot Logbook Results Config About 1 Signal selection 2 Current Y Axis 17 04 22 0 C 3 Signal Units
31. modify and store methods It also shows all method parameters and a graphical view of the timetable settings In opposite to the edit options from the status tab which reflects an online editing mode the Method tab allows offline editing and storing Changes that are made are not directly applied to a currently running fractionation If you want to permanently change a method you work in the method tab offline mode and save a modified default method under a different file name Method selection Status Method Piot Logbook Results Config About Parameter per step Run um Timtable graphic Parameter in Timetable Addition of Timetable lines LEN Volt Hour Voltage Current Power Time kVh v uA mW h mm Delete Save Save As ce ci A Q N Save Delete options Timetable 0 00 2 00 4 00 6 00 8 00 co MEN me Set Aid Delete Figure 14 Editing Methods 46 OFFGEL User Manual Operation 5 In a pull down menu for Method Selection you can select methods that are present on the internal drive of the instrument or on the USB stick inserted at the rear of the instrument These methods can either be default methods which were supplied with the instrument or user modified methods A 3100 OFFGEL fractionators method generally consist out of two method steps which are the focusing and the hold phase The methods do not contain the platform temperature as parameter During the focusing step the sample fractionation is t
32. on a separate worksheet TI_56 CSV 10 4 2006 16 30 57 DE00000064 Tray File name Run at Instrument Serial Number with Tray 2000 o pe ES E eo t ino PES 3 1000 Curb i 0 2 4 5 8 10 12 4 16 Time h Figure 22 Plotted graph of run parameters y axes versus time h 60 OFFGEL User Manual OFFGEL User Manual Operation 5 If a worksheet is manually deleted or the data file is reloaded the corresponding graph is deleted If a plot to a certain worksheet is repeatedly generated the previous plot version is deleted Table 5 gives details of the different run parameters Table5 Run parameters Run Parameter Description VoltTT PowerTT kVHours Volt Power Temp Top Cover CurrA CurrB CurrC UAhA pARB pARC Fixed settings of Volt and Power of the method that was used kVHours Volt Power measured during the fractionation for the complete tray Temperature of the cooling platform during the fractionation The events or time intervals when the top cover was opened during a fractionation are marked with the red diamond symbol on the X axis If there are no Top Cover events registered the Top Cover check box is automatically unchecked Currents measured for individual tray lanes A H Measured uA hours acquired for individual tray lanes A H 61 4 instru ient Configuration Instrument Configuration Figure 23 The Configuration tab
33. the resident system then restart After the restart it will update or downgrade the main system and restart again Please delete the folders for update or downgrading the firmware if you wish to leave the USB stick in the instrument The presence of these folders decreases the boot up speed of the instrument at future starts OFFGEL User Manual Agilent 3100 OFFGEL Fractionator User Manual 7 Troubleshooting Troubleshooting the Instrument 92 Troubleshooting the Electrodes 100 Troubleshooting the LAN Connection 101 Troubleshooting the Application 102 This chapter provides support for troubleshooting the instrument and the application RE Agilent Technologies 91 7 Troubleshooting Troubleshooting the Instrument 92 Status Indicators Status LED The instrument has a status LED on the front top right position see Figure 2 on page 17 With the power switched on the LED can have the following states Table 8 LED status overview LED Status Description Comment Green The instrument is performing a fractionation Yellow The instrument is currently not ready Arunning fractionation will be paused Possible reasons if the top cover is opened the set temperature of the cooling platform has not been reached top cover is open Red The instrument has detected an error A running fractionation will be aborted see Table on page 94 Off The instrument is not performing a fractionation The set tempera
34. tion with the furnishing use or per formance of this document or of any information contained herein Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 oras a commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 June 1987 or any equivalent agency regulation or contract clause Use duplication or disclo sure of Software is subject to Agilent Tech nologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 June 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 June 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in any technical data Safety Notices CAUTION A CAUTION notice de
35. to a USB Memory stick inserted at the rear of the instrument can be done Status Method Plot Logbook Results Config About Results Calendar ko mm Macro Download Deine o March 2010 o Result files Download USB export selection Su Mo Tu We Th Fr Sa Excel 2003 USB export progress Excel 2007 Desult files 16 15 1 00244 16 15 1 00245 16 16 00246 Copying files to USB drive Export to USB Day Month Year Figure 17 Results from fractionations In the Results Calendar you can select the month and day when results were created Clicking onto a date populates the files available for this selection You can download the Agilent 3100 OFFGEL Fractionator plotter for data analysis by Microsoft Excel at any date containing data files Under Data 54 OFFGEL User Manual Operation 5 analysis is a link for Macro Download of the two versions for Microsoft Office 2003 or 2007 available Next to this you find the links for a Result file download Under operating system Windows 7 with Internet Explorer 8 a download of result file is done best by a right click onto the respective download link and selection of the option Save Target As A double click will open the result file which is comma separated values for electrical data in an Internet Explorer window It is advantageous to save a file first open and analyze the result file by using the macro from within Microsoft Excel see Chapter 7 on page 105 The download
36. with 3100 OFFGEL Low Res Kit pH 3 10 12 well frames OR for use with 3100 OFFGEL Low Res Kit pH 4 7 12 well frame OFFGEL default method for peptides for use with 3100 OFFGEL Low Res Kit pH 3 10 12 well frames OR for use with 3100 OFFGEL Low Res Kit pH 4 7 12 well frame IPG IEF in gel default method for protein and peptides for use with 24 cm IPG strips pH 3 10 OR pH 4 7 IPG IEF in gel default method for protein and peptides for use with 13 cm IPG strips pH 3 10 OR pH 4 7 use for hardware diagnosis as described in Checking High Voltage Electrodes and Electrode Connectors on page 97 65 5 Operation Please note that latest OFFGEL methods do have a voltage limit of 4500 V This voltage limit reduces the risk of arcing and damage to the tray in case of dried out electrode pads Preparing a Tray for the OFFGEL Mode To prepare a tray for the OFFGEL mode refer to the Kit Manual 5969 1582 for detailed instructions The manual for OFFGEL fractionation is included in every consumable kit see Table 1 on page 19 Preparing a Tray for the Standard IPG IEF Mode You can use any IPG strip for fractionations in the standard IPG IEF mode in gel mode Follow the recommendations of the IPG strip supplier about rehydration loading of the sample to the IPG strip The tray of the Agilent 3100 OFFGEL Fractionator system can be used as a rehydration tray The following workflow uses an IPG strip already loaded with sample and
37. Agilent 3100 OFFGEL Fractionator User Manual E Agilent Technologies Notices Agilent Technologies Inc 2010 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Manual Part Number 63100 90001 Edition 04 2010 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn Microsoft is a U S registered trademark of Microsoft Corporation Software Revision This guide is valid for B 06 30 or higher revi sions of the Agilent 3100 OFFGEL Fraction ator Firmware where the last two digits of the Firmware revision refer to minor revi sions of the Firmware that do not affect the technical accuracy of this guide Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connec
38. EL High Res Kit pH 4 7 5188 6426 3100 OFFGEL Low Res Kit pH 4 7 5188 6427 3100 OFFGEL Starter Kit 5188 6444 OFFGEL Protein Test Sample 5188 6428 OFFGEL User Manual 19 20 Product overview Instrument and Accessories OFFGEL User Manual Agilent 3100 OFFGEL Fractionator User Manual 2 Site requirements and specifications Power Considerations 22 Bench Space 22 Environment 22 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC 23 Sound Emission 23 Instruction for Use 24 Physical Specifications of the Agilent 3100 OFFGEL Fractionator 25 This chapter describes information about site requirements that have to be met so that the Agilent 3100 OFFGEL Fractionator performs within specifications RE Agilent Technologies 21 2 Site requirements and specifications Site Requirements for the Agilent 3100 OFFGEL Fractionator Power Considerations CAUTION The Agilent 3100 OFFGEL Fractionator power supply has wide ranging capabilities and accepts any line voltage in the range 100 240 V with a line frequency of 50 60 Hz see also on page 25 for more details Consequently there is no voltage selector in the rear of the instrument Shock hazard or damage to your Agilent 3100 OFFGEL Fractionator may result if the Agilent 3100 OFFGEL Fractionator is connected to a line voltage higher than specified Bench Space The Agilent 3100 OFFGEL Fractionator requires sufficient desk or laboratory ben
39. Electrode Assy G3100 60002 includes 1 tray 1 fixed electrode and 1 movable electrode USB Memory Stick Kit G4208 68700 maximum capacity is 2GB 12 Well Frame Set 5188 8012 Set includes 12 well frames 12 wells cover seals electrode pads glycerol solution cover fluid and manual To be used with 13 cm IPG gel strips and respective ampholytes for use with 5188 6444 5188 6425 5188 6427 24 Well Frame Set 5188 8013 Set includes 12 well frames 24 wells cover seals electrode pads glycerol solution cover fluid and manual To be used with 24 cm IPG gel strips and respective ampholytes for use with 5188 6444 5188 6424 5188 6426 156 OFFGEL User Manual Index Symbols 126 A 120 accessories 33 add instrument 82 anode 19 anode connector 18 cleaning 72 aviso 148 baudrate 78 bench space 22 browser interface 28 C cables de alimentaci n 148 CAN connection 34 cathode 19 cathode connector 18 caution 114 120 126 136 142 cleaning 70 anode connector 72 cooling platform 72 electrodes 70 top cover 75 tray 71 clock 42 COM ports 78 compact flash card 34 35 compatibilidad electromagn tica 149 condensation 22 154 configuration switch 29 34 63 101 configure directories 81 firmware update tool 80 connection CAN 34 LAN 34 USB 34 OFFGEL User Manual Consignes de s curit 136 consumable kits 19 155 cont
40. Manual 117 8 Safety Safety Information 118 OFFGEL User Manual Agilent 3100 OFFGEL e 00 e O e e e i 120 Ne RE Agilent Technologies OX TIL Agilent 3100 OFFGEL sE H p 119 Agilent 3100 OFFGEL NN Bra o RE he HITS RNCS
41. Produktion und geplante Nutzung des Ger ts Agilent Technologies bernimmt keine Haftung im Falle der Missachtung dieser Voraussetzungen durch den Kunden Ein Warnhinweis weist Sie auf Situationen hin die Personensch den oder eine Zerst rung der Ausr stung verursachen k nnen bergehen Sie nicht diesen Hinweis bevor Sie die Warnung vollst ndig verstanden haben und die entsprechenden Bedingungen erf llt sind ACHTUNG Ein Achtungs Hinweis weist auf eine Situation hin die zu einer Besch digung des Agilent 5100 ALP oder einem Datenverlust f hren kann bergehen Sie nicht diesen Achtungs Hinweis bevor Sie ihn vollst ndig verstanden haben und die entsprechenden Bedingungen erf llt sind 130 Ger tesicherheit Dies ist ein Ger t der Sicherheitsklasse mit einem Anschluss f r Schutzerde und wurde gem internationalen Sicherheitsstandards hergestellt und getestet OFFGEL Benutzerhandbuch Sicherheit Netzkabel Fur den Agilent 3100 OFFGEL Fractionator werden unterschiedliche Netzkabel als Optionen angeboten Die Steckbuchse ist bei allen Kabeln identisch und wird in die Netzanschlussdose auf der R ckseite des Rechners gesteckt Der Stecker ist bei jedem Kabel unterschiedlich und entspricht den Netzsteckdosen des betreffenden Landes WARNUNG Betreiben Sie Ihr Ger t niemals an einem Netzanschluss ohne Erdung Benutzen Sie niemals ein anderes Netzkabel als das von Agilent Technologies f r Ihren Bereich d
42. This default IP is hard coded in the instrument and enabled by configuration dip switches on the backside of the instrument Figure 7 on page 34 For assignment of an IP address to the PC follow these steps 1 Plug Cross Over LAN cable into the LAN connector of the PC 2 Make sure the second end of this Cross Over LAN cable is accessible to be inserted to the 3100 OFFGEL fractionator Place the PC at a suitable location and power it on Log on with an user account that has administrator rights Click on Start and select Control panel ao OT a c Go to the Section Network and Internet and select the item View network status and tasks 7 Select and click with the right mouse button onto the item Change adapter settings OFFGEL User Manual Installation 4 8 At the item Local Area Connection click with the right mouse button onto Properties 9 Within the Properties select Internet Protocol Version 4 10 Select Use the following IP address 11 Enter the following information IP Adress 192 168 254 10 Subnet Mask 255 255 255 0 12 Select Ok to finish 13 Close remaining windows You may now proceed switching on the Agilent 3100 OFFGEL Fractionator and open the Internet Explorer on the control PC Type the IP address 192 168 254 11 to view the web based user interface See Chapter 5 The User Interface starting on page 40 for the description of the interface A short description on how to set up the instrument and compute
43. Y Axis 4 Graph area 5 Display options 15 30 15 40 15 50 16 00 16 10 16 20 16 30 16 40 Unzoom Stacked Tray Figure 15 Plot of Signals OFFGEL User Manual 49 5 50 Operation Zoom in Plots Using a PC with a mouse connected the mouse wheel can be used for zooming Both axes scales can be changed independent from each other The mouse wheel input will change Y Scale as long as the mouse pointer is located nearby the Y Axis inside the graph area If the mouse pointer is anywhere else within the graph area the X Scale will be changed Alternatively the arrow buttons in combination with control button from the control PC keyboard can be used for zooming in the plot area Left arrow together with control button allows zooming in X axis right arrow together with control button allows zooming out X axis up arrow together with control button allows zooming in Y axis down arrow together with control button allows zooming out the Y axis The Y axis is zoomed starting in the middle of the plot and the X axis from the left of the plot During zooming with arrows the cursor must be set over the graph area The minimum time scale span in zoomed views is 20 minutes The unzoom function resets the plot scaling to the initial values which are defined by the button overlay stacked for the Y axis and a 2 hours time frame for the X Axis Using the the iPod the pinch gesture is used to zoom the plot area Pan Drag in Plots On the PC t
44. able address can be edited by clicking into the fields adjacent to the IP Address Subnet Mask and Default Gateway A click onto the button Set stores the entered values to the connected Agilent 3100 OFFGEL Fractionator A Default LAN Configuration with default address settings which cannot be changed by the user is present in the lower panel of the configuration tab Please see instructions for first time connection of the Agilent 3100 OFFGEL Fractionators to an instrument on page 31 Chapter 4 Please note that any configuration change will be active only after an instrument restart On the Configuration tab you can also trigger module identification A click onto the button Identify sends a command to the connected Agilent 3100 OFFGEL Fractionator to let the control LED flash for a few seconds in all three colors OFFGEL User Manual 63 Additional Information Additional Information The About page provides information on the revision of the currently active firmware of the connected instrument It provides links to helpful Agilent web pages such as the Agilent 3100 OFFGEL Fractionator for latest news on the instrument or to a download location for the firmware update tool Here you find copyright information of the Webinterface and the used open source Javascript libraries For service purposes the diagnosis buffer can be downloaded here Provide the diagnosis buffer together with result files to you local Agilent customer care center
45. ablished parameters for TCP IP communication are used Typically speed and duplex mode are determined by auto negotiation When using the RS 232 cable the Firmware Update Tool uses the following port settings when performing the update Baudrate 19200 overrides the PC settings No Parity 8 Data Bits e 1 Stop Bit COMI COM2 etc The settings are set automatically for the time of the firmware update OFFGEL User Manual Maintenance 6 Note that the upgrade process with LAN will take proximately 5 minutes An update with RS 232 cable connection can take up to 2 hours No fractionations can be performed during this time Setup of the Firmware Update Tool 1 Start the Firmware Update Tool via the shortcut see Figure 27 The following initial screen is displayed NE File Action Options Help Menu All Instruments Buttons for quick access of functions All Instruments Instrument window shows system and modules No instruments configured Instrument name and communication type information Module details Figure 28 Start up screen When you move with the cursor over a information is given about its function Alternatively you may use the menu items File Open Protocol File Exit Action Connect Disconnect Select for update Identify module Start Cancel update OFFGEL User Manual 79 6 Maintenance Options Configure directories Add Delete
46. aking place typically at higher voltages until a given Volt Hour value kVh is reached or a given Time h mm has elapsed The hold step is typically done at lower voltages after the fractionation just to maintain the achieved separation until the fractions are recovered from the wells The Volt Hour value kVh or Time h mm are limiting parameters for the duration of the fractionation The default methods have values that were found to be suitable for a broad range of applications and samples The parameters Voltage Current and Power are by Ohms Law in a correlation for a given fractionation in an individual strip lane The Method parameters Voltage Current and Power are limiting factors during a run One of the parameters typically current is at the limit Please save modifications before changing to another tab Changes will be lost otherwise The focus step of a method can be sub devided into different time based steps if desired Create a time based program using the lower part of the method tab A Timetable for the parameters Voltage Current and Power is visualized in a graphical way with color coding per value In case time table entries are done the parameters set above are used as starting values at time 0 00 Select the desired parameter for example voltage in the timetable pane from a drop down menu Set the value in the adjacent field Changing from one value setting to another one at a second timepoint can be done as one step
47. aminated with mineral oil Mineral oil tends to creep across the electrodes and tray wells once they are moistened However if the electrodes do not dry out during fractionation this does not impair the focusing results carefully pipette the mineral oil and avoid spillages when inserting the tray into the instrument make sure that no oil is spilled out of tray Insert the tray before adding the mineral oil to the instrument OFFGEL User Manual Looking OFFGEL User Manual Troubleshooting 7 at Run Parameters The results of all run parameters can be exported as a csv file and thus can be visualized with Microsoft Excel Agilent Technologies has designed the Agilent 3100 OFFGEL fractionator plotter which is a Excel add on and facilitates the analysis of the run parameters see How to Display Exported Results on page 58 The following examples show typical results for good runs as well as some impaired results where the root cause of the problem is known Example 1 Typical Results of the Protein Test Sample The fractionation of the protein test sample is a way to check that the Agilent 3100 OFFGEL Fractionator in combination with the consumables kit is working in your hands and functions as a system checkout test You find detailed information on how to run the protein test sample in the kit manual Figure 45 shows a typical result for the electrical run parameters of the protein test sample using t
48. anni alle persone o alla strumentazione Non procedere oltre se non si certi di aver compreso le istruzioni e soddisfatto le condizioni indicate AVVERTENZA Un segnale di avvertimento richiama l attenzione su una situazione che protrebbe compromettere la funzionalit o le prestazioni del sistema Frazionatore OFFGEL Agilent 3100 e causare perdite di dati Non procedere oltre se le situazioni indicate non sono state risolte 142 Sicurezza dello strumento Lo strumento appartiene alla Classe di sicurezza dotato di terminale di protezione con messa a terra ed stato prodotto e collaudato secondo standard di sicurezza internazionali Manuale per l utente del frazionatore OFFGEL Sicurezza Cavo di alimentazione ATTENZIONE Il sistema Frazionatore OFFGEL Agilent 3100 puo essere utilizzato con vari tipi di cavi di alimentazione acquistabili a parte Il lato con il connettore femmina identico su tutti i tipi di cavi e deve essere collegato alla presa di alimentazione sul retro dello strumento Il lato con il connettore maschio varia a seconda dei cavi e dei tipi di prese utilizzati nei vari paesi Non collegare lo strumento ad una presa di corrente sprovvista di messa a terra Non utilizzare un cavo di alimentazione diverso da quello fornito da Agilent Technologies per l uso nel proprio paese ATTENZIONE Per un funzionamento ottimale e per rispettare le normative di sicurezza EMC importante utilizzare solo i cavi
49. anual Contents Regular Cleaning Procedure 70 Cleaning Procedure for Heavily Contaminated Electrodes 70 Cleaning the Trays 71 Cleaning the Cooling Platform 72 Cleaning the Anode Connector 72 Cleaning the Top Cover and the Housing of the Instrument 75 Firmware Update 76 Installation of the Firmware Update Tool 77 How to un install the Firmware Update Tool 77 Installation of the Firmware on yourPC 77 Connect your PC to the Agilent 3100 OFFGEL Fractionator 78 Setup of the Firmware Update Tool 79 Configure the Firmware Update Tool 80 Doing the Firmware Update 84 The Log file 87 Firmware Change via USB Stick 88 Steps for Firmware Change via USB 89 Troubleshooting 91 Troubleshooting the Instrument 92 Status Indicators 92 Status LED 92 Status Bars 93 Error Messages 93 Hardware Diagnosis 96 Checking the LED andthe cover sensor 96 Checking Temperature Control ofthe Cooling Platform 96 Checking High Voltage Electrodes and Electrode Connectors 97 Troubleshooting the Electrodes 100 Contents 10 Checking Fixed and Movable Electrodes 100 Troubleshooting the LAN Connection 102 Looking at Run Parameters 105 Troubleshooting the Application 101 Example 1 Typical Results of the Protein Test Sample 105 Example 2 Current Drop During Fractionation 107 Example 3 Reduced Electrical Charge in One Lane 108 Safety 113 Safety Information 114 Power Cords 114 Operation 115 Safety Symbols 116 Chemical and Biolog
50. ate T ool Firmware Update Tool A Firmware Update Help Update Tool s Firmware Update Report Firmware Figure 27 Icon on your desktop and the Program Folder How to un install the Firmware Update Tool The Firmware Update Tool has an un install routine If required remove the Firmware Update Tool via the Uninstall Firmware Update Tool function within the program folder see Figure 27 Installation of the Firmware on your PC 1 Download the Agilent 3100 OFFGEL Fractionator firmware from the Agilent web site OFFGEL User Manual 77 6 78 Maintenance http www chem agilent com scripts DownloadsUtilities asp onto your PC 2 Double click the exe file to extract the content into a folder default is c Program Files Agilent LC Firmware Update Tool The Firmware Update Tool can be customized to use different folders as firmware location Please change the path under Options Configure Directories from within the Firmware Update Tool Connect your PC to the Agilent 3100 OFFGEL Fractionator Your PC has to be connected by a Cross Over LAN cable or an normal LAN cable to the Agilent 3100 OFFGEL Fractionator Please see instructions for Installation in Chapter 4 In case you would like to use the alternative approach to connect the PC with the Firmware update tool by RS232 see a description of connections at the rear of the instrument see Figure 7 on page 34 When using a LAN connection to the instrument the est
51. ch space dimensions and weights are shown in Table on page 25 Approximately 15 cm 6 inches of clear space at the rear of the instrument for electric connections is needed Environment CAUTION The Agilent 3100 OFFGEL Fractionator will work within the specifications of ambient temperatures and relative humidity described in Table 2 on page 25 Do not store ship or use the Agilent 3100 OFFGEL Fractionator under conditions where temperature fluctuations could cause condensation within the Agilent 3100 OFFGEL Fractionator Condensation will damage the electronics If the Agilent 3100 OFFGEL Fractionator was shipped in cold weather leave it in its box and allow it to warm up slowly to room temperature to avoid condensation 22 OFFGEL User Manual Site requirements and specifications 2 Put the Agilent 3100 OFFGEL Fractionator only on a flat and horizontal work top Inclinations of more than 0 5 will cause the instrument to malfunction The Agilent 3100 OFFGEL Fractionator is designed to operate in a controlled electromagnetic environment EN61326 A1 where RF transmitters such as mobile phones should not be used in close proximity The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC adopted by EU Commission on 13 February 2003 is introduced producer responsibility for all Electric and Electronic appliances from 13 August 2005
52. ction in Chapter 4 Once the connection is reestablished review and correct the IP address in the Config tab of the web interface Call Agilent Technologies if the problem cannot be solved OFFGEL User Manual 101 7 Troubleshooting Troubleshooting the Application 102 Impaired focusing results can have many different root causes In addition to hardware errors the impaired focusing results can be sample and or handling related Therefore always make sure that you set up your focusing experiment exactly as described in the kit manual The sample concentration should be between 50 ug ml and 5 mg ml and the salt content of your sample buffer should be at most 10 mM Run only samples similar in protein content and buffer composition on the same tray otherwise it is highly recommended that you use separate trays With every focusing a results file containing all electrical parameters during a run are stored to the compact flash card and can be exported The traces also provide useful information for troubleshooting see How to Display Exported Results on page 58 and some examples are discussed in this chapter see Looking at Run Parameters on page 105 The following table gives an overview of symptoms you might experience when performing OFFGEL experiments their possible causes and actions required for remedy OFFGEL User Manual Table 12 Troubleshooting the OFFGEL application Troubleshooting 7 Symptom Pos
53. ctionator RE Agilent Technologies 27 3 PC Browser and LAN Requirements PC Browser and LAN Requirements Browser Interface The Agilent 3100 OFFGEL Fractionator can be controlled from an internet browser interface The communication of browser application and the instrument is based on the TCP IP network protocol as it is used for display of webpages from the world wide web There is no specific control module on the instrument or dedicated software necessary on the computer Table 3 lists browsers that were in principle capable for control of the instrument There are no dedicated restrictions on the operating system Table3 Examples of Compatible Browsers Type Internet Explorer IE 8 0 Microsoft recommended Internet Explorer IE 7 0 Microsoft Safari 4 0 4 Apple Firefox 3 5 Mozilla The Internet Explorer IE 6 0 Microsoft is not suitable for controlling the 3100 OFFGEL fractionator due to missing functionality in the browser 28 OFFGEL User Manual PC Browser and LAN Requirements 3 LAN The 3100 OFFGEL Fractionator comes with a default IP address 192 168 254 11 that cannot be changed The instrument can also be assigned a user settable static IP address This custom IP address can be set after an initial connection via the default address Configuration dip switches at the instrument rear allow to select which of the two addresses are used by the 3100 OFFGEL Fractionator This configuration is applied whe
54. d nicht als biologisch wirksame Dichtung f r die R ckhaltung von Aerosolen und Fl ssigkeiten ausgef hrt Handhaben Sie K rperfl ssigkeiten mit Vorsicht da sie Krankheiten bertragen k nnen Kein bekannter Test bietet vollst ndige Sicherheit daf r dass sie frei von Mikroorganismen sind Einige der ansteckendsten Hepatitis B und C und HIV 1 V Viren atypische Mykobakterien und bestimmte systemische Pilze machen die Notwendigkeit f r Schutz vor Aerosolen noch wichtiger Befolgen Sie immer die rtlichen Bundes und Landesvorschriften f r die Handhabung biologisch gef hrlicher Stoffe bei der Entsorgung von gef hrlichem biologischen Abfall z B kontaminierte Well Rahmen Handhaben Sie alle infekti sen Proben entsprechend guter Laborpraktiken und methoden um die Ausbreitung von Krankheiten zu vermeiden Entsorgen Sie alle Losungsmittelabfalle und Abfallprodukte gem den entsprechenden Gesundheits und Sicherheitsvorschriften OFFGEL Benutzerhandbuch 133 Sicherheit Sicherheitshinweise 134 OFFGEL Benutzerhandbuch Fractionneur Agilent 3100 OFFGEL M Manuel de l utilisateur ee e O 090 e Securite i Consignes de s curit 136 Ce chapitre contient des informations de s curit importantes concernant le Fractionneur Agilent 3100 OFFGEL RE Agilent Technologies 135 S curit Consignes de s curit Les consignes g n rales de s curit suivantes doivent tre observ es lors
55. e display of webpage content retrieved from the instrument Please see Chapter 4 for establishing the LAN connection Switch your instrument on and type its IP address default IP address 192 168 254 11 into the address line of your browser to display the user interface The user interface provides several tabs to control the instrument see also Figure 9 on page 41 Status which is also the application start screen shows the current state of both trays You can select methods to be used start and stop processing and modify used parameters interactively in this view Method screen is used to view modify and store methods It shows all method parameters and a graphical view of the timetable settings Plot screen allows to view actual over time in a XY plot You can select up to 4 signals The XY plot allows to zoom in and out and to pan the plot area Logbook shows a time ordered list of the occurred events since last start of the instrument Result view allows to download result files from fractionations using the browser download function and also to copy result files of a time span to a connected USB Memory stick inserted at the rear of the instrument Config screen allows to setup view LAN configuration You can trigger module identification by flashing the control LED at the connected Agilent 3100 OFFGEL Fractionator About page shows the revision information of the firmware and links to helpful Agilent web pages Here you fi
56. e pared de cada pa s o regi n Manual de usuario de OFFGEL Seguridad No utilice nunca el instrumento en una toma de alimentacion que no est conectada AVISO M SE a tierra No utilice nunca un cable de alimentacion que no haya sido suministrado por Agilent Technologies y dise ado para su zona geogr fica AVISO No utilice nunca cables que no hayan sido suministrados por Agilent Technologies para asegurar un correcto funcionamiento asi como el cumplimiento de las normativas de seguridad o de compatibilidad electromagn tica Operacion Antes de conectar el instrumento a la alimentaci n el ctrica debe seguir las instrucciones de instalaci n para hacer funcionar el instrumento con seguridad No retirar las cubiertas del instrumento mientras est funcionando Antes de encender el instrumento todos los cables de tierra alargadores auto transformadores y aparatos conectados al mismo deben conectarse a tierra mediante un enchufe adecuado Si se interrumpe la conexi n a tierra podrian producirse lesiones graves Siempre que se sospeche que la conexi n a tierra se ha interrumpido debe bloquearse el aparato para evitar cualquier manipulaci n Evitar todo mantenimiento del instrumento mientras est bajo tensi n No se deben sustituir los componentes del instrumento mientras est encendido No manejar el instrumento en presencia de gases o vapores inflamables El funcionamiento de cualquier instrumento el ctric
57. ecurity settings under tools gt macro gt security allow macros to be run The following button is generated in your Microsoft Excel 2003 tool bar or is available under Add Ins Custom Toolbars for Microsoft Excel 2007 Figure 19 Agilent 3100 OFFGEL Fractionator toolbar button OFFGEL User Manual Operation 5 3 Click on the Agilent 3100 OFFGEL Fractionator toolbar button to activate the following user interface Agilent 3100 OFFGEL Fractionator X Make Graphic List Box lt A lt I lt I Uncheck all Figure 20 Agilent 3100 OFFGEL Fractionator user interface You can close the Agilent 3100 OFFGEL Fractionator user interface by clicking the close window button You can reopen it by clicking the Agilent 3100 OFFGEL Fractionator toolbar button 4 Click Open File select an exported file with the browser and open the file The file must be an unchanged Agilent 3100 OFFGEL Fractionator csv file 5 The file name of the loaded file appears in the Agilent 3100 OFFGEL Fractionator list box Figure 21 Multiple files can be loaded simultaneously You can toggle between them by clicking on the file name OFFGEL User Manual 59 5 Operation Reviewing Result Files Agilent 3100 OFFGEL Fra Figure 21 Agilent 3100 OFFGEL Fractionator user interface result file opened 6 Select all run parameters which should be plotted 7 Click Make Graphic The plot is displayed
58. eklarierte Netzkabel WARNUNG Benutzen Sie keine anderen als die von Agilent Technologies gelieferten Kabel um eine einwandfreie Funktion und Ubereinstimmung mit den gesetzlichen Vorschriften sicher zu stellen Betrieb Befolgen Sie die Installationsanweisungen bevor Sie das Gerat an die Netzspannung anschlie en Zur Gew hrleistung eines sicheren Betriebes Entfernen Sie w hrend des Betriebs keine Ger teabdeckung Vor dem Einschalten des Ger ts m ssen alle Erdungsanschl sse Verl ngerungskabel Autotransformato ren und angeschlossene Ger te an eine geerdete Steckdose angeschlossen werden Jede Unterbrechung der Schutzerdung bedeutet ein potenzielles Stromschlagrisiko das zu schweren Verletzungen f hren kann Wenn davon ausgegangen werden muss dass der Schutz beeintr chtigt wurde muss das Ger t ausgeschaltet und gegen unbeabsichtigten Betrieb gesichert werden F hren Sie keine Wartungsarbeiten am Ger t unter Spannung aus Ersetzen Sie keine Komponenten w hrend das Ger t eingeschaltet ist Betreiben Sie das Ger t nicht in Gegenwart leicht entz ndlicher Gase oder D mpfe Der Betrieb elektrischer Ger te in einer solchen Umgebung stellt ein hohes Sicherheitsrisiko dar Bauen Sie nur Originalteile ein und nehmen Sie keine nicht autorisierten Anderungen am Ger t vor OFFGEL Benutzerhandbuch 131 Sicherheit Sicherheitssymbole Tabelle 13 Sicherheitssymbole am Ger t und in diesem Handbuch Symbol Besc
59. ent it is the responsibility of the user to ensure that all necessary safety regulations guidelines precautions and practices are adhered to accordingly This includes also the handling of genetically modified organisms Ask your laboratory safety officer to advise you about the level of containment required for your application and about proper decontamination or sterilization procedures to follow if fluids escape from containers Observe all cautionary information printed on the original solution containers prior to their use Because leaks spills or loss of sample may generate aerosols observe proper safety precautions Agilent 3100 OFFGEL Fractionator covers have not been designed as bioseals for aerosol or liquid containment Handle body fluids with care because they can transmit disease No known test offers complete assurance that they are free of micro organisms Some of the most virulent Hepatitis B and C and HIV I V viruses atypical mycobacteria and certain systemic fungi further emphasize the need for aerosol protection Always follow local state and federal biohazard handling regulation when disposing of biohazardous waste material e g contaminated compartment frames Handle all infectious samples according to good laboratory procedures and methods to prevent spread of disease Dispose of all waste solutions and products according to appropriate environmental health and safety guidelines OFFGEL User
60. equently to be the stop button A fractionation will start only if the instrument is ready for example if the platform temperature for the given method is reached Readiness is given by the status indicator An idle instrument will start immediately the OFFGEL User Manual 43 5 44 Operation 4 tal otatus tab fractionation Not ready indication on for example tray temperature will lead to a waiting step An open lid will cause the not ready information and application of the voltage is done only if the lid is finally closed see also Table 8 on page 92 The active Run status for each tray is additionally indicated by a blue tray corona around the respective tray shaped interface An Agilent 3100 OFFGEL Fractionators method generally consist out of two method steps which are the focusing and the hold phase Methods are described in more detail under Working with Methods on page 46 The run parameter shown in the status screen are the specified limits for volt hours time voltage power and current of the loaded method The bar graph on the left side visualizes the actual values online The bar size is in relation to the set point in addition the numeric value is displayed by a tool tip if the mouse pointer is place over the respective bar The bar graph labeled with letters from a to h on the right side visualizes the current measured as actual per lane for each strip online Click into the lane area to swap from displaying C
61. er elements are out of limits Check that the top cover closes tightly The cover sensor might be defective Call Agilent Technologies The temperature sensor is defective Call Agilent Technologies The fan might be defective Call Agilent Technologies The leak sensor is defective Call Agilent Technologies Check for condensation or dirt on anode connector electrodes or tray Clean the the parts as described in Cleaning the Agilent 3100 OFFGEL Fractionator on page 70 The high voltage module might be defective Call Agilent Technologies A Peltier element might be defective Call Agilent Technologies In case you wish to call Agilent Service for support on the 3100 OFFGEL fractionator provide a detailed error description exported results from your latest sample runs diagnosis buffers content of the instruments log book and firmware revision 95 7 96 Troubleshooting Hardware Diagnosis With the following few steps you can verify if your instrument is performing to its specifications Note that a meaningful hardware diagnosis requires a fully functional communication to the instrument Checking the LED and the cover sensor 1 Switch on or restart the instrument and watch the Status LED during startup The Status LED must show all three LED colors for approximately one second during startup in the following order yellow green red If this is not the case a LED is broken and needs repa
62. er que toutes les r gles de s curit directives pr cautions et pratiques sont respect es en cons quence Ceci concerne galement la manipulation des organismes g n tiquement modifi s Demandez au responsable de la s curit de votre laboratoire de vous conseiller en ce qui concerne le niveau de confinement requis par votre application et les proc dures de d contamination ou de st rilisation suivre en cas de fuites de fluides Avant toute utilisation de solutions respectez toutes les pr cautions indiqu es sur leur emballage d origine Les fuites coulements ou pertes d chantillon pouvant g n rer des a rosols observez les r gles de s curit ad quates Les capots du Fractionneur Agilent 3100 OFFGEL n ont pas t concus pour assurer l tanch it aux a rosols et aux liquides Manipulez les fluides corporels avec pr caution ceux ci tant susceptibles de transmettre des maladies Aucun test connu ne peut garantir qu ils sont exempts de micro organismes Certains des plus virulents tels que les virus de l h patite B et C et le HIV I V les mycobact ries atypiques et certains champignons syst miques exigent une protection renforc e contre les a rosols Suivez toujours les r gles nationales et locales de manipulation lors de la mise au rebut de mat riel usag risque biologique par exemple des cadres compartiments contamin s Manipulez tous les chantillons infectieux selon les proc dure
63. es 69 6 Maintenance Cleaning the Agilent 3100 OFFGEL Fractionator Materials used in the instrument that may come in contact with liquid reagents are resistant to chemicals typically used for isoelectric focusing IEF Always turn off the instrument and unplug the power cord before doing any cleaning procedures Always wear powder free gloves when handling parts that are in contact with sample or mineral oil CAUTION Do not use ketones such as acetone or strong acids to clean the instrument Cleaning the Electrodes 70 Electrodes can be used multiple times They are designed to be robust but might suffer from permanent coating of the surface and from arcing events during fractionations Regular Cleaning Procedure Both the fixed and the movable electrodes have to be cleaned before every use 1 Wipe the electrode pins with water using a lint free tissue 2 Wipe the electrode pins with isopropanol using a lint free tissue to remove the mineral oil 3 Thoroughly air dry the electrodes or dry well with a lint free tissue prior to use Cleaning Procedure for Heavily Contaminated Electrodes If for any reason not only the electrode pins but the complete electrodes are contaminated use the following cleaning procedure OFFGEL User Manual CAUTION Maintenance 6 Rinse the complete electrodes with hand warm tap water Do not exceed a water temperature of 50 C Use a mild detergent such as dishwashing detergen
64. eshooting the Application TI_53 csv 8 29 2006 10 59 53 DE00000034 Tray 1400 800 Volt VoltTT uAh gt 400 Time h Figure 48 Example 3 one lane shows reduced electrical charge The root cause of this failure was a conductivity problem in lane F OFFGEL User Manual 109 7 Troubleshooting Troubleshooting the Application 110 OFFGEL User Manual Agilent 3100 OFFGEL Fractionator e User Manual al e a e e 000 ee 8 e Safety e e e e Safety Information 114 This chapter contains safety information relevant to the Agilent 3100 OFFGEL Fractionator RE Agilent Technologies 113 8 Safety Safety Information The following general safety precautions must be observed during all phases of operation service and repair of the Agilent 3100 OFFGEL Fractionator All safety instructions should be read and understood before installation operation and maintenance of the instrument Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design manufacture and intended use of the instrument Agilent Technologies assumes no liability for the customer s failure to comply with these requirements A warning alerts you to a situation that could cause physical injury or damage to the equipment Do not proceed beyond a warning until you have fully understood and met the indicated conditions CAUTION A caution alerts you to situations
65. et 1 5 mm 8710 0641 4 Compact Flash Card 1819 0277 5 Cross over LAN cable 2023 0203 6 USB Memory Stick Kit G4208 68700 Part numbers are of April 2010 and might be subject of changes OFFGEL User Manual 33 4 Setting up the Agilent 3100 OFFGEL Fractionator 34 After you have unpacked the Agilent 3100 OFFGEL Fractionator and positioned it in a suitable place on your bench you need to do the following Install the Compact Flash Card Insert the USB memory stick Connect the power cord The Agilent 3100 OFFGEL Fractionator has the following connectors at the rear Configuration CAN RS232 LAN Compact Line power Dip Switch not used not used Flash Card USB Figure 7 Connections at the rear of the instrument 1 Insert the compact flash card into the dedicated position 2 Connect the Cross Over LAN cable to the LAN connector Make sure the second end is available to be connected to the control PC at a later point in time OFFGEL User Manual OFFGEL User Manual Installation 4 Setting up the Agilent 3100 OFFGEL Fractionator Be sure to not to confuse the LAN connector with the CAN connector The web browser access to the instrument will not work when connecting it to the CAN connector 3 Connect the power cable to the instrument and to the line power 4 Verify configuration dip switches Figure 8 Switches N 1 to 6 must be down Switches N 7 and 8 must be up 5 Switch on the instrumen
66. f the Agilent 3100 OFFGEL Fractionator Installation This chapter describes how to install the Agilent 3100 OFFGEL Fractionator Operation This chapter details how to operate the Agilent 3100 OFFGEL Fractionator It familiarizes the user with all control functions from the internet browser interface It describes the necessary steps to perform a fractionation and set up a LAN connection to a PC Maintenance This chapter describes cleaning procedures that should be used to do maintenance on your instrument at regular intervals or before every use In addition it provides instructions on how update the firmware Troubleshooting This chapter provides support for troubleshooting the instrument and the application Safety This chapter contains safety information relevant to the Agilent 3100 OFFGEL Fractionator Appendix This chapter contains details on non condensing condition as well as ordering information for consumables and spare parts OFFGEL User Manual Related Documents The Agilent 3100 OFFGEL Fractionator can be used with different consumable kits which each are supplied with a Kit Manual 5969 1582 Kit Manual OFFGEL User Manual OFFGEL User Manual Contents Contents 1 Productoverview 13 Intended Use 14 Fractionation Principles ofthe Agilent 3100 OFFGEL Fractionator 14 Features 16 Instrument and Accessories 17 2 Site requirements and specifications 21 Site Requirements for the Agile
67. f the firmware update process This file is useful as a compliance document to prove that the module firmware files were updated successfully or as diagnostics information in the case of problems see The Log file on page 87 Therefore name the protocol file appropriately 2 Press OK OFFGEL User Manual 81 6 Maintenance Firmware Update 82 3 Select the Add Instrument button L RES Update Tool File Action Options Help EFA ak O sl s Figure 31 Select the Add Instrument button 4 Enter a name for the system e g System 1 5 Enter the TCP IP address information in the instrument address field e g 168 192 254 11 for the default address of the instrument Set here the static IP address of your instrument in case you connected it to a network In case you connect alternatively with RS232 cable enter the valid COM port connection information in the instrument address field e g COMI Add Instrument Select an instrument name address from the lists below or edit the text fields directly Instrument Name 3100 OFFGEL 1 Instrument Address fi 34 40 27 54 cars Figure 32 Add instrument screen OFFGEL User Manual Maintenance 6 Update 6 Press OK Firmware Update Tool File Action Options Help cm All Instruments 3100 OFFGEL 1 All Instruments Instrument Address 3100 OFFGEL 1 134 40 27 54 Figure 33 Add instrument screen 7 Double click on All I
68. f the fixed electrode anode connector OFFGEL User Manual 103 7 104 Troubleshooting Table 12 Troubleshooting the OFFGEL application continued Symptom Possible Cause Action 3 the corresponding sample has higher salt concentration or higher protein peptide content than the other samples Run only samples similar in protein content and buffer composition on the same tray After fractionation has finished the external wells of the frame have reduced liquid or are empty During IEF a water transport is observed due to migrating ions that drag the water in their hydration shell to the electrodes Furthermore water is dragged from wells at the basic strip end cathodic end of strips pH 3 10 to wells in the neutral pH range due to the ionization of DTT Therefore after OFFGEL electrophoresis the wells close to the electrodes are sometimes filled with less liquid than the inner wells or are even empty If low recovery in the extreme wells is a problem refill the corresponding wells with diluted OFFGEL Stock Solution without ampholytes 4 parts OFFGEL Stock Solution without ampholytes plus one part water one hour prior to the end of the focusing phase Sparking at the electrodes short circuits at the electrodes make sure that the electrodes and the electrode connectors are completely dry After fractionation has finished the cooling platform is cont
69. he 3100 OFFGEL Low Res Kit pH 3 10 The method used for this fractionation was the default method OG12PR00 which has a current limit of 50 uA a voltage limit of 8000 V a power limit of 200 mW and a volt hour limit of 20 kVh With this setup the fractionation is running in current limiting mode Therefore the current signal in Figure 45 is stable and at 50 LA Please note that latest OFFGEL methods do have a voltage limit of 4500 V This is reduce the risk of arcing and damage to the tray in case of dried out electrode pads 105 7 Troubleshooting Troubleshooting the Application TI_56 CSV 10 4 2006 16 30 57 DE00000064 Tray 2000 a 8 x 9 i vot un E 60 KVous i 1000 re Cun i current electrical power Volt kVhours charge Figure 45 Example 1 typical results for the Protein test sample using the 3100 OFFGEL Low Res Kit pH 3 10 The run stopped when the volt hour limit of 20 kVh was reached Both volt signal and power signal are increasing throughout the run since the sample is depleted of ions and thus the resistance is increasing After fractionation the protein test samples should be focused in fraction 3 see Figure 46 106 OFFGEL User Manual Troubleshooting kDa TT band e m in wo nr co oo a 8 E E E E E c c c c a e S S S S s S s 3 EFT Ei S E E E E E E E E 95 63 ss 4 NM 23 NEN is E 65 m 35 A U
70. he graph area is panned by pressing the left mouse button while the mouse is located in the plot area moving the mouse and releasing it Panning is performed separate for X and Y direction depending on which movement is larger Using the iPod the panning is performed by a two finger drag within the plot area OFFGEL User Manual Operation 5 Switching the Y Scale in Plots Printing OFFGEL User Manual The source for the Y Axis scaling can be changed from one to another signal by clicking onto the colored line of the signal in the graph area The color of the y Axis will change to reflect the color of the signal it is related to Once a signal is loaded freshly the Y axis will use this as source for scaling The stacked and overlay view in the plots specifies the arrangement of the signals on the plot pane Stacked display allows visualization of signals with four individual offsets on the axis while overlay has the same offset for all signals Signals can mask each other when overlayed of Plots Printing of signal plots is typically possible and is based on the browser features For printing of signal plots select the print button of the browser or select the Print dialog under the browser file menu Selected elements of the OFFGEL interface such as the graph of the selected signals will be printed 51 Instrument Logbook Instrument Logbook The Logbook shows events occurred on the instrument The list consists of all event
71. hreibung Das Ger t ist mit diesem Symbol gekennzeichnet wenn der Bediener A das Benutzerhandbuch lesen sollte um Verletzungen zu vermeiden und das Ger t vor Sch den zu sch tzen Chemische und biologische Sicherheit WARNUNG Beachten Sie beim Arbeiten mit L sungsmitteln die entsprechenden Sicherheitsma nahmen z B Tragen von Schutzbrille Schutzhandschuhen und Schutzbekleidung gem der Beschreibung im Datenblatt zur Materialhandhabung und Sicherheit des Losungsmittelherstellers insbesondere bei Verwendung toxischer oder gef hrlicher L sungsmittel 132 OFFGEL Benutzerhandbuch Sicherheit WARNUNG Wenn pathogene toxische oder radioaktive Proben in diesem Ger t verwendet werden sollen ist der Bediener verantwortlich f r die Einhaltung aller notwendigen Sicherheitsrichtlinien Vorsichtma nahmen und Praktiken Dies betrifft auch die Handhabung genetisch modifizierter Organismen Fragen Sie den Sicherheitsbeauftragten Ihres Labors nach den notwendigen Sicherheitsma nahmen f r Ihre Anwendung und den korrekten Dekontaminierungs und Sterilisierungsverfahren f r den Fall dass Fl ssigkeit aus Beh ltern entweicht Lesen Sie vor der Verwendung alle Vorsichtshinweise auf den Originall sungsmittelbeh ltern Da durch Lecks verschiittetes L sungsmittel und Probenverluste Aerosole entstehen k nnen beachten Sie die entsprechenden Sicherheitsma nahmen Die Abdeckungen des Agilent 3100 OFFGEL Fractionator sin
72. i n manejo y mantenimiento del instrumento Si no se cumplen estas normas o los avisos espec ficos que aparecen en diversas partes de este manual se invalidan los est ndares de seguridad de dise o fabricaci n y utilizaci n de este instrumento Agilent Technologies no se responsabiliza del incumplimiento por parte del usuario de estas normas Un aviso advierte sobre una situaci n que podr a derivar en lesiones personales AVISO o da os al equipo No continuar tras un aviso hasta haber entendido y cumplido totalmente las condiciones indicadas 7 Una precauci n advierte de situaciones que podr an limitar la funcionalidad o el EHECAUCION rendimiento del Fraccionador 3100 OFFGEL de Agilent y dar lugar a una p rdida de datos No continuar tras un mensaje de precauci n hasta haber tomado en consideraci n plenamente las condiciones indicadas Seguridad del instrumento Este es un instrumento de seguridad de Primera Clase dotado de un terminal de toma de tierra y ha sido fabricado y comprobado de acuerdo a las normas internacionales de seguridad Cables de alimentaci n 148 Con el Fraccionador 3100 OFFGEL de Agilent se ofrecen diferentes cables de alimentaci n opcionales El extremo hembra de todos ellos es id ntico Este encaja en la entrada de alimentaci n de la parte trasera del instrumento El extremo macho de todos los cables de alimentacion es diferente y esta dise ado para adaptarse a los enchufes d
73. ical Safety 119 120 121 121 122 125 126 126 127 127 128 Sicherheit 129 Sicherheitshinweise 130 116 122 OFFGEL User Manual Contents Netzkabel 131 Betrieb 131 Sicherheitssymbole 132 Chemische und biologische Sicherheit 132 8 S curit 135 Consignes de s curit 136 Cordons secteur 137 Utilisation 137 Symboles de s curit 138 S curit chimique et biologique 138 8 Sicurezza 141 Informazioni per la sicurezza 142 Cavo dialimentazione 143 Funzionamento 143 Simboli di sicurezza 144 Sicurezza chimica e biologica 144 8 Seguridad 147 Informaci n de seguridad 148 Cables de alimentaci n 148 Operaci n 149 S mbolos de seguridad 150 Seguridad qu mica y biol gica 150 9 Appendix 153 Non condensing Conditions 154 Ordering Information 155 List of Consumables 155 List of Spare Parts 156 OFFGEL User Manual 11 Contents 12 OFFGEL User Manual Agilent 3100 OFFGEL Fractionator M User Manual e O e 1 e i Product overview e Intended Use 14 Features 16 Instrument and Accessories 17 This chapter gives an overview of the Agilent 3100 OFFGEL Fractionator It describes the intended use and the fractionation principles of the instrument highlights its featu
74. igh voltage power supplies which provide voltages to the two trays On the left side of the cooling platform is the anode connector across the cooling platform are two cathode connectors see Figure 3 Figure 3 View inside the instrument Attached to each tray are a fixed electrode and a movable electrode that establish the connection to the anode and the cathode connectors see Figure 4 Each tray accommodate up to eight samples Each OFFGEL fractionation requires a frame to build up the well compartments as depicted schematically in Figure 1 There are two different frame lengths available depending on the consumable kit and application chosen The 12 well frame is typically used for fractionations requiring lower resolution the 24 well frame is used for fractionations requiring higher resolution Trays and electrodes can be ordered as spare parts if needed for ordering details refer to Table 15 on page 156 OFFGEL User Manual Product overview 1 Instrument and Accessories Movable Electrode Cathode Frames Fixed Electrode Anode Tray Fixed Electrode Anode Movable Electrode Cathode Tray Cover Seals Figure 4 Electrodes trays and frames The frames are included in the consumable kits The available consumable kits are listed in Table 1 Table 1 Consumable kits Product Order Number 3100 OFFGEL High Res Kit pH 3 10 5188 6424 3100 OFFGEL Low Res Kit pH 3 10 5188 6425 3100 OFFG
75. ilent Technologies 31 4 Installation Unpacking the Agilent 3100 OFFGEL Fractionator Unpacking the Agilent 3100 OFFGEL Fractionator Before you begin the installation ensure that the place you have chosen meets the site requirements as specified in Site Requirements for the Agilent 3100 OFFGEL Fractionator on page 22 The shipment consists of three boxes one containing the computer optional one containing the instrument and one containing the power cable 1 Unpack the computer Please proceed with starting and configuring the computer as described the documentation associated with it Make sure to retain the administrative rights until the system setup is complete 2 Openthe box in which the Agilent 3100 OFFGEL Fractionator was shipped On the top you will find a box containing the accessories and a Declaration of Conformity Figure 5 Opened instrument shipment box 3 Take out the accessory box and lift the instrument with its foam packing out of the outer box 32 OFFGEL User Manual Installation 4 TI n lrzme ha Anilant 21N0 NEENE Era n Unpacking the Agilent 3100 OFFGEL Frat tor Figure 6 Instrument inner shipment box 4 Remove the foam packing and the inner box and place the instrument in a suitable place 5 Check that you have the complete set of accessories available using Table 4 Table4 Accessories Description Part Number 1 User Manual G3100 90001 2 Quick Installation guide G3100 90111 3 Hex key s
76. imientos adecuados de descontaminaci n o esterilizaci n que deben seguirse si se producen escapes de l quidos de los recipientes Deben tenerse en cuenta toda la informaci n de aviso impresa en los recipientes originales de las soluciones con anterioridad a su uso Puesto que las fugas vertidos o p rdidas de muestras pueden generar aerosoles deben tenerse en cuenta las precauciones de seguridad adecuadas Las cubiertas del Fraccionador 3100 OFFGEL de Agilent no se han dise ado como cierres biol gicos hermeticos para la contenci n de aerosoles o l quidos Los fluidos corporales deben manipularse con cuidado ya que pueden transmitir enfermedades Ninguna prueba conocida ofrece una certeza total de que est n exentos de microorganismos Algunos de los mas virulentos como los virus de la hepatitis B y C y el HIV I V microbacterias atipicas y determinados hongos sist micos ponen de relieve la necesidad de una protecci n contra los aerosoles Siempre se deben observar los reglamentos oportunos en materia de manipulacion de sustancias biol gicas peligrosas al desechar residuos biol gicos de alto riesgo ej estructuras de compartimento contaminadas Todas las muestras infecciosas se deben manipular de acuerdo con los procedimientos y m todos de laboratorio adecuados con el fin de evitar la propagacion de enfermedades Todos los residuos de soluciones y productos deben desecharse de acuerdo con directrices adecuadas de segu
77. ing condition the humidity has to be below the plotted curves OFFGEL User Manual Ordering Information List of Consumables Appendix 9 Table 14 Agilent 3100 OFFGEL Fractionator consumable kits Product Order Number Product Description 3100 OFFGEL 3 10 Low Res Kit 3100 OFFGEL 3 10 High Res Kit 3100 OFFGEL 4 7 Low Res Kit 3100 OFFGEL 4 7 High Res Kit 3100 OFFGEL Starter Kit OFFGEL Protein test sample 5188 6425 5188 6424 5188 6427 5188 6426 5188 6444 5188 6428 includes frames 12 well IPG strips pH 3 10 13 cm cover seals and fractionation reagents for 12 samples includes frames 24 well IPG strips pH 3 10 24 cm cover seals and fractionation reagents for 12 samples includes frames 12 well IPG strips pH 4 7 13 cm cover seals and fractionation reagents for 12 samples includes frames 24 well IPG strips pH 4 7 24 cm cover seals and fractionation reagents for 12 samples includes frames 6x 12 well 6x 24 well IPG strips pH 3 10 3x 13 cm 3x 24cm IPG strips pH 4 7 3x 13 cm 3x 24cm cover seals and fractionation reagents for 12 samples includes beta lactoglobulin as a protein test sample enough for more than 100 tests OFFGEL User Manual 155 9 Appendix List of Spare Parts Table 15 Agilent 3100 OFFGEL Fractionator spare parts Product Order Number Product Description Tray Set G3100 60007 includes 4 replacement trays
78. ir Call Agilent Technologies for further help Otherwise proceed with step 2 2 Set the cooling platform temperature to 20 C and wait until the cooling platform temperature is reached and the LED is turned off 3 Open the top cover The LED must turn yellow The interface must show Cover Open in the status bar This confirms that the cover sensor is working properly A failed cover sensor would be recognized by the instrument and result in an error see Table on page 94 Checking Temperature Control of the Cooling Platform For this test the ambient temperature must be within specification maximum 30 C if the cooling platform is set to 20 C and conditions should be non condensing 1 Set the cooling platform temperature to 20 C and wait until the cooling platform temperature is reached and the LED is turned off 2 Set the cooling platform temperature to 30 C and measure the time until the LED is turned off The temperature ramp up should not take longer than 5 minutes 3 Set the cooling platform temperature back to 20 C and measure the time until the LED is turned off The temperature ramp down should not take longer than 15 minutes OFFGEL User Manual Troubleshooting 7 Checking High Voltage Electrodes and Electrode Connectors 1 Make sure your two electrode sets and trays are completely dry and clean 2 Assemble both trays with the electrodes without frames and insert them to the instrument Attach the mo
79. malattie Non esistono test in grado di garantire che siano completamente privi di microrganismi La presenza di virus mortali come epatite B e C ed HIV I V microbatteri atipici ed alcuni funghi sistemici aumenta ulteriormente la necessit di protezione dagli aerosol Attenersi sempre alle normative in vigore localmente quando si devono smaltire rifiuti contenenti materiali biologicamente pericolosi ad esempio telai di comparti contaminati Maneggiare i campioni infetti secondo la buona prassi di laboratorio ed utilizzare metodi adatti per prevenire la diffusione della malattia Smaltire tutte le soluzioni ed i prodotti di scarico in base alle linee guida per la sicurezza e la salute ambientale Manuale per l utente del frazionatore OFFGEL 145 146 Sicurezza Informazioni per la sicurezza Manuale per l utente del frazionatore OFFGEL Fraccionador 3100 OFFGEL de Agilent M Manual de usuario 00 Q 0 e Seguridad Informaci n de seguridad 148 En este cap tulo encontrar informaci n de seguridad importante relativa al Fraccionador 3100 OFFGEL de Agilent RE Agilent Technologies 147 Seguridad Informacion de seguridad Las siguientes precauciones generales de seguridad deben observarse durante todas las fases de funcionamiento mantenimiento o reparaci n del Fraccionador 3100 OFFGEL de Agilent Deben leerse y comprenderse perfectamente todas las instrucciones de seguridad antes de la instalac
80. method or save the method once the fractionation is finished Wait until the fractionation is finished to delete this method Table 11 Error messages hardware related LED is red Error message Comment Action Cooling platform temperature The cooling platform temperature has not been error EE 20302 reached A leak was detected in the module Use the logbook to determine date and time the fault occurred EE 00064 Make sure the ambient temperature and the set cooling platform temperature are within instrument specifications A Peltier element might be defective Call Agilent Technologies A leak has been detected by Check if there is liquid the leak sensor spillage or condensation on the cooling platform and near the leak sensor and wipe it away For details refer to Cleaning the Cooling Platform on page 72 Acknowledge the error in the logbook to reset the red LED status OFFGEL User Manual OFFGEL User Manual Table 11 Troubleshooting 7 Error messages hardware related LED is red continued Error message Comment Action Cover sensor failed EE 20300 Opening and closing the top cover is no longer recognized Temperature sensor failed EE 20301 Fan failed EE 00068 Leak EE 00080 EE 00081 EE 00082 or EE 00083 High voltage EE 20305 The voltage did not reach the set point Peltier Error EE 20304 The voltage or currents of the Pelti
81. n the instrument is powered on the next time A detailed description on the steps for instrument setup is given under Chapter 4 Installation starting on page 31 The instrument is not enabled for DHCP Dynamic Host Configuration Protocol Therefore an automated assignment of network configurations from a server is not possible For incorporation into a local network a static IP address is needed Tested Configurations The configuration of the typical bundle equipment Part Number G3100AA of the system is described below It was successfully tested and consists of a3100 OFFGEL Fractionator a Netbook HP Mini 5102 Operating system Windows 7 Starter Edition Internet Explorer 8 0 aconnection by a Cross Over LAN cable Another successfully tested configuration consisted of e a3100 OFFGEL Fractionator with wired LAN connection to a local Network Apple iPod Touch 8G FW Revision 3 1 2 including a Safari Browser with wireless LAN access to the local Network OFFGEL User Manual 29 3 30 PC Browser and LAN Requirements PC Browser and LAN Requirements OFFGEL User Manual Agilent 3100 OFFGEL Fractionator M User Manual 000 ee 4 270 Installation i Unpacking the Agilent 3100 OFFGEL Fractionator 32 Setting up the Control PC 36 Setting up the Agilent 3100 OFFGEL Fractionator 34 This chapter describes how to install the Agilent 3100 OFFGEL Fractionator and set up a LAN connection to a PC RE Ag
82. nator Table 2 Physical Specifications Type Specification Comment Weight 13 kg 29 Ibs Dimensions 157 x 355 x 427 mm Height x width x depth Line voltage 100 240 VAC Wide ranging capability Line frequency 50 60 Hz Power consumption 140 VA Ambient operating 5 40 C 41 104 F see also humidity temperature specification Ambient non operating 40 70 C 40 158 F Humidity 0 90 temperature Humidity 15 80 at 5 40 C 41 104 F see details in non condensing e g max humidity is Appendix 9 55 96 if the cooling platform is used Non condensing 10 C below ambient temperature Conditions Cooling platform temperature 10 35 C 50 95 F maximal 10 C below ambient temperature High voltage power supply 500 V 10 kV maximal 150 pA per gel strip maximal 1 W per gel strip Operating altitude 2000 m 6666 ft Non operating altitude 4500 m 15000 ft Safety Standards IEC CSA UL Installation Category Il Safety Class Pollution Degree 2 OFFGEL User Manual 25 2 Site requirements and specifications Site Requirements for the Agilent 3100 OFFGEL Fractionator 26 OFFGEL User Manual Agilent 3100 OFFGEL Fractionator M User Manual 000 ee 3 e PC Browser and LAN Requirements e x Browser Interface 28 LAN 29 Tested Configurations 29 This chapter describes information about requirements for your PC browser and LAN that have to be met for the control of the Agilent 3100 OFFGEL Fra
83. nd copyright information of the Webinterface and the used open source Javascript libraries You can click on the tabs in order to see the different screens When data such as method parameters were not save they might disappear upon leaving the tab OFFGEL User Manual Operation 5 The User Interface Web Browser Tab OFFGEL interface tabs Instrument clock amp sob E Asien offo ron X Aglent 3100 OFFGEL Fracti N ethod Plot Logbook Results Config About 1250 Idle 220 C Tray OG12PE01 v Volt Hours 20kVh Time 100 00 hm Platform temperature Status indicator USB indicator Application Lock A Ei Tray method Method edit 0 Fractionation start Voltage 4500 V Power 200 mW Current 50 uA A co 0 N OO Gl F 65 N O O Edit Start Tray Il OG24PE01 v Volt Hours 50 kVh Time 100 00 hm Voltage 4500V Power 200 mw Current 50 uA I CIN Edit Start Figure 9 Overview on Agilent 3100 OFFGEL Fractionator interface OFFGEL User Manual 41 5 Operation General Functions few elements of the screen shown in Figure 9 on page 41 are common to many tabs Besides the Status indicator and the USB indicator this are the Instrument Clock and the Platform Temperature You can adjust these parameters by clicking onto this area The dialog shown in figure 5 appears and will allow you to set the date time and the platform temperature DEG 25 2 2010 Kol sc 1 on Temperature a
84. notes a haz ard It calls attention to an operat ing procedure practice or the like that if not correctly performed or adhered to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated conditions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indicated condi tions are fully understood and met OFFGEL User Manual In This Guide This book describes how to operate the Agilent 3100 OFFGEL Fractionator including the browser based user interface 3100 OFFGEL Fractionator Control PC Chapter Overview 1 Product overview This chapter gives an overview of the Agilent 3100 OFFGEL Fractionator It describes the intended use and the fractionation principles of the instrument highlights its features and outlines the different instrument components 2 Site requirements and specifications This chapter describes information about site requirements that have to be met so that the Agilent 3100 OFFGEL Fractionator performs within specifications OFFGEL User Manual 3 PC Browser and LAN Requirements This chapter describes information about requirements for your PC browser and LAN that have to be met for the control o
85. nstruments to expand the view M dfyouhave a control module G3101A connected in parallel and want to update the Control Module select Options and check Support Control Module The control module is optional and might come from earlier instrument configurations 8 Select the instruments name e g OFFGEL 1 and click on the Connect to Instrument button Firmware Update Tool File Action Options Help Instrument Address 4 Figure 34 Select 3100 OFFGEL 1 All modules within OFFGEL 1 are listed with serial number firmware revision and build number G3100A refers to the Agilent 3100 OFFGEL Fractionator instrument In case of an additionally connected control module this will show up as G3101A OFFGEL User Manual 83 6 Maintenance Firmware Update Firmware Update Tool BAE File Action Options Help All Instruments 3100 OFFGEL 1 gt G31004 DE00000070 Instrument noFfeEL Address Revision Build G31004 DE00000070 B 06 30 0002 Figure35 Modules of 3100 OFFGEL 1 are listed Doing the Firmware Update The following describes the general workflow for a firmware update of the 3100 OFFGEL fractionator instrument 1 Select a module Firmware Update Tool DOOR File Action Options Help All Instruments 3100 OFFGEL 1 Current Firmware Yuste Information Revision History lt G31004 DE00000070 Identity Module Q Identity Main System Resident System
86. nt 3100 OFFGEL Fractionator 22 Power Considerations 22 Bench Space 22 Environment 22 The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC 23 Do not dispose of in domestic household waste 23 Sound Emission 23 Instruction for Use 24 Physical Specifications of the Agilent 3100 OFFGEL Fractionator 25 3 PC Browser and LAN Requirements 27 PC Browser and LAN Requirements 28 Browser Interface 28 LAN 29 Tested Configurations 29 4 Installation 31 Unpacking the Agilent 3100 OFFGEL Fractionator 32 Setting up the Agilent 3100 OFFGEL Fractionator 34 Setting up the Control PC 36 OFFGEL User Manual 7 Contents 5 Assignment of an IP Address to the Control Computer Local Network Connection optional 37 Operation 39 The User Interface 40 General Functions 42 Status tab 43 Working with Methods 46 Displaying Data Graphically 49 Zoom in Plots 50 Pan Drag in Plots 50 Switching the Y Scale in Plots 51 Printing of Plots 51 Instrument Logbook 52 Exporting Results 54 Reviewing Result Files 57 How to find the exported files on the USB stick 57 How to Display Exported Results 58 Instrument Configuration 62 Additional Information 64 Performing a Fractionation 65 Preparing a Tray forthe OFFGEL Mode 66 Preparing a Tray for the Standard IPG IEF Mode 66 StartingaRun 68 Maintenance 69 Cleaning the Agilent 3100 OFFGEL Fractionator 70 Cleaning the Electrodes 70 36 OFFGEL User Manual OFFGEL User M
87. o en estas circunstancias constituye un riesgo para la seguridad No instalar componentes de repuesto que no correspondan al instrumento ni realizar modificaciones no autorizadas Manual de usuario de OFFGEL 149 Seguridad Simbolos de seguridad Tabla 13 S mbolos de seguridad empleados en los instrumentos y en los manuales Simbolo Descripci n El instrumento aparece marcado con este s mbolo para indicar que el usuario debe consultar el Manual de usuario con el fin de evitar posibles lesiones al operador y para proteger el equipo contra posibles da os Seguridad qu mica y biol gica AVISO Cuando se trabaje con disolventes debe seguir los procedimientos de seguridad apropiados por ejemplo gafas guantes y ropa adecuada descritos en las especificaciones sobre el tratamiento de material y datos de seguridad que suministra el proveedor especialmente cuando se utilicen productos toxicos 0 peligrosos 150 Manual de usuario de OFFGEL AVISO Seguridad Si se tiene previsto utilizar muestras pat genas t xicas o radioactivas en este instrumento es responsabilidad del usuario asegurar que se observen adecuadamente todos los reglamentos directrices precauciones y pr cticas de seguridad Esto tambi n incluye la manipulaci n de organismos modificados gen ticamente Solicitar asesoramiento al responsable de seguridad del laboratorio acerca del nivel de contenci n necesario para cada aplicaci n y sobre los proced
88. of a fractionation OFFGEL User Manual 57 5 58 How to Display Exported Results The results of all run parameters can be exported as comma separated value csv file and thus can be visualized with Microsoft Excel Agilent Technologies has designed the Agilent 3100 OFFGEL fractionator plotter which is an Excel add on and facilitates the analysis of the run parameters The tool is downloadable from the instrument in revisions for Microsoft Excel 2003 and 2007 The tool is also provided via the Agilent Technologies web page http www agilent com Life Sciences and Chemical Analysis Solutions Technical Support Downloads amp Utilities or direct via http www chem agilent com scripts DownloadsUtilities asp and can be downloaded free of charge Note that the tool was designed for Microsoft Excel 2003 and 2007 with US American system local settings only and has not been tested with other versions of Microsoft Excel However it is expected that the tool also runs under international versions and international system local settings 1 In Microsoft Excel 2003 open the downloaded file Agilent 3100 OFFGEL Fractionator xla You may also want to copy the file in the XLSTART folder for office 2003 it can be C Program Files Microsoft Office OFFICE11 XLSTART so that the program is loaded automatically when Excel is started 2 Click Enable Macros when the security warning window pops up If you cannot run the program check that Excel s
89. oling Once the cooling platform Wait Temp Il platform has not been reached temperature is reached the however a fractionation was already fractionation will be started started on one of the trays automatically Error Messages OFFGEL User Manual Table 10 Error messages due to wrong operation Error message Solution USB drive full Exchange USB drive or delete Exchange the USB drive or delete files on the files USB drive File export failed The selected files cannot get Exchange the USB drive or delete files on the copied to USB drive Check if drive is full USB drive 93 7 94 Troubleshooting Table 10 Error messages due to wrong operation continued Error message Solution Result Storage Empty No result data to copy CF Card is empty or not present No USB drive USB drive not present or incompatible drive format Cannot overwrite default method A default method cannot be overwritten Change method name The method file lt name gt on lt drive gt cannot be overwritten It is currently running on lt tray gt Delete Denied The method lt name gt from lt drive gt is currently in use on lt tray gt Insert a compact flash card for details refer to Figure 7 on page 34 Insert a USB drive Make sure the format is compatible for details refer to page 54 Change the method name to save modification of default methods Change the method name to save modification to this
90. option avec le Fractionneur Agilent 3100 OFFGEL L extr mit femelle est identique pour tous les cordons secteur Elle se branche dans la prise secteur pr vue cet effet l arri re de l instrument Le c t male de chaque cordon secteur est sp cifique au pays ou la r gion de destination Ne jamais faire fonctionner un instrument si la borne de terre de l alimentation secteur n est pas effectivement reli e la terre Ne jamais utiliser un cordon secteur diff rent de celui sp cifiquement fourni par Agilent Technologies pour la r gion d utilisation Afin d assurer un fonctionnement correct et conforme la r glementation sur la compatibilit lectromagn tique ne jamais utiliser un cordon secteur diff rent de celui sp cifiquement fourni par Agilent Technologies Utilisation Avant la mise sous tension suivez les instructions d installation pour utiliser l instrument en toute s curit Ne pas retirer les capots de l instrument pendant son fonctionnement Avant de mettre l instrument sous tension toutes les bornes de mise la terre les rallon ges les auto transformateurs et les p riph riques qui y sont connect s doivent tre branch s sur une prise munie d une fiche de terre de protection Toute interruption de la ligne de terre constitue un risque d lectrocution pouvant engendrer des blessures corporelles graves S il s av re que la protection n est plus assur e l utilisation de l instrument devra
91. or applied as a linear gradient The point in time when this change should happen during the method execution can be given Press the Add button to add more method steps You also can edit any time table entry by highlighting it By selecting one line the corresponding values get populated in the parameter line After modifying values you can apply them to the method with the Set button The platform temperature is no method parameter and cannot be changed in the timetable For changing platform temperature please see page 42 OFFGEL User Manual 47 5 48 Operation Printing of methods is typically possible and is based on the browser features Select the print button of the browser or select the print dialog under the browser file menu Selected elements of the OFFGEL method such as the values for focussing and hold phase the graphical display of the timetable and the time table will be printed Please note that Voltage setting beyond 4500 kV may lead to increased arcing under certain experimental conditions Such conditions are dried pads which can cause burnt spots in the tray Default methods are therefore limited to 4500 kV Functions in the method pane are Delete Removes the method from the instrument Only user defined methods can be deleted Save Stores the shown setpoints in the currently selected method Save As Opens a file dialog that shows the currently existing methods and allows the user to specify a new
92. quency 22 25 line voltage 22 25 loading capacity 16 102 lock instrument 45 log book 52 log file 87 M method DIAGNOSE 97 movable electrode 19 network 28 34 36 62 non condensing conditions 25 96 154 non operating altitude 25 non operating temperature 25 0 OFFGEL mode 14 66 operating altitude 25 operating temperature 25 operation 115 121 127 131 137 143 149 P pH resolution 16 physical specifications 25 pl 14 platform temperature 41 plotter graph 60 toolbar button 58 user interface 59 power considerations 22 power consumption 25 power cords 114 121 126 137 143 power outlet 115 121 127 137 143 power supply 22 precaucion 148 precauciones de seguridad 148 OFFGEL User Manual protein test sample 19 105 107 155 protocol file 88 report screen 87 result 58 105 RF transmitters 23 riesgo para la seguridad 149 RS 232 baudrate 78 connection 34 run parameter review 105 S safety hazard 115 121 127 137 143 safety information 114 safety instructions 114 120 126 136 142 safety precautions 114 120 126 136 142 safety standards IEC CSA UL 25 safety symbols 116 122 127 132 138 144 sample concentration 16 102 serial number 17 setup firmware update tool 79 instrument 34 Sicherheitshinweise 130 sound emission 23 spare parts 156 start arun 68 status bar titles 93 status LED 17 status overview
93. r is given in a Quick Guide G3100 90111 shipped with the instrument Do connect the PC only to the 3100 OFFGEL fractionator Connecting the PC with this manually set IP address to your local area network will most likely create network issues Local Network Connection optional For connecting the 3100 OFFGEL fractionator to a local network request a static IP address subnet mask and gateway adress from your network administrator Although it is intended to use the 3100 OFFGEL fractionator in a local network at a later point in time you have to access the instrument first with the default IP address 192 168 254 11 The steps for this are given under Assignment of an IP Address to the Control Computer on page 36 and Setting up the Agilent 3100 OFFGEL Fractionator on page 34 When a local LAN connection via Cross Over LAN cable between control PC and 3100 OFFGEL fractionator is established you can set the provided values for static IP address subnet mask and gateway address under Config of the OFFGEL User Manual 37 4 38 Installation web based User Interface Please see the description of the web based User Interface in Chapter 5 The User Interface starting on page 40 and find addition information on the Instrument Configuration on page 62 After you have set the custom IP address subnet mask and gateway address do the following steps 1 Power the instrument off 2 Install a conventional LAN cable from
94. res and outlines the different instrument components RE Agilent Technologies 13 1 Product overview Intended Use 14 The Agilent 3100 OFFGEL Fractionator system fractionates proteins or peptides based on their isoelectric points pl in liquid phase OFFGEL mode In addition the system performs standard in gel isoelectric focusing IEF using immobilized pH gradient gel IPG strips in gel mode The system consists of an instrument that can be controlled via LAN by a User Interface based on a typical web browser Earlier instruments were controlled by a handheld control module which can still be used in parallel The web interface provides complete control and monitoring of the Agilent 3100 OFFGEL Fractionator The system is capable of running a total of 16 samples in parallel 2x8 OFFGEL pre fractionation of complex protein and peptide sample provides additional sensitivity of downstream analysis tools such as LC MS In addition the pl is a useful parameter that can be used for validation of peptide MS hits Fractionation Principles of the Agilent 3100 OFFGEL Fractionator The Agilent 3100 OFFGEL Fractionator performs isoelectric focusing of proteins or peptides in immobilized pH gradient IPG gel strips The 3100 s OFFGEL electrophoresis differs from conventional gel electrophoresis in that the sample components do not remain in the gel Instead they are recovered from a buffer solution making the recovery much easier than
95. ridad e higiene medioambientales Manual de usuario de OFFGEL 151 152 Seguridad Informacion de seguridad Manual de usuario de OFFGEL Agilent 3100 OFFGEL Fractionator M User Manual 000 ee 9 e Appendix Non condensing Conditions 154 Ordering Information 155 This chapter contains details on non condensing conditions as well as ordering information for consumables and spare parts RE Agilent Technologies 153 9 Appendix Non condensing Conditions 154 The Agilent 3100 OFFGEL Fractionator is specified to be operated only under non condensing conditions and with a cooling platform temperature of maximum 10 C below ambient temperature For example assuming a typically experimental setup of 20 C on the cooling platform and an ambient temperature of 25 C the humidity must be below 73 A plot of humidity versus ambient temperature for the cooling platform at 20 C 25 C and 30 C is depicted in Figure 1 For non condensing conditions the humidity has to be below the plotted curves Maximum Humidity vs Ambient Temperature 100 95 90 85 80 75 Humidity 70 65 60 55 50 20 21 22 23 24 25 26 27 2 29 30 31 12 33 34 35 36 37 38 39 40 Ambient Temperature PC Cooling Platform 20 C Cooling Platform 25 C Cooling Platform 30 C Figure 1 Plot of humidity versus ambient temperature assuming the cooling platform to be set at 20 C 25 C or 30 C For non condens
96. rol 83 control module 17 83 control PC 36 cooling platform 18 cleaning 72 temperature 96 cover seals 19 csv file 58 105 D date 42 diagnosis hardware 96 diagnostic buffer 64 dimensions 25 E electrode assy 156 electrodes cleaning 70 troubleshooting 100 electromagnetic environment 23 EMC regulations 115 121 127 137 143 environment 22 error messages 94 Excel macro 58 export 54 F features 16 firmware doing the update 84 log file 88 revision 64 update tool 77 update with USB 88 fixed electrode 19 flammable gases 121 127 137 143 fractionation principle 15 frame 19 fumes 121 127 137 143 Index G gases inflamables 149 general functions 42 hardware diagnosis 96 humidity 25 154 Informazioni per la sicurezza 142 in gel mode 14 16 66 71 installation firmware update tool 77 instrument 34 instrucciones de seguridad 148 instrument accessories 33 anode connector 18 cathode connector 18 configuration 62 cooling platform 18 errors 94 firmware update 76 hardware diagnosis 96 installation 34 serial number 17 setup 34 spare parts 156 status LED 17 92 top cover 17 troubleshooting 92 Instrument lock 45 IP adress 29 36 62 78 101 IPG IEF mode 66 IPG strips 14 IPG strip 66 isoelectric points 14 157 L LAN connection 28 34 63 78 101 firmware update tool installation 77 firmware update tool setup 79 line fre
97. rrent pattern for all eight lanes Figure 44 a Status Method Plot Logbook Results Config About 17 22 Run 20 0 C Tray 2 DIAGNOSE v e Volt Hours 2 5 kVh Time 0 15 h m Voltage Power Current OO Edit Stop Figure 44 Typical current pattern on tray when every lane is filled with water 13 Press Stop on both trays to stop the runs EET T If you require further assistance call Agilent Technologies and send the corresponding ue results file of this diagnose test to the customer call center together with a detailed description exported results from your latest sample runs diagnosis buffers content of the instruments log book and firmware revision OFFGEL User Manual 99 7 Troubleshooting Troubleshooting the Electrodes 100 Electrodes can be used multiple times They are designed to be robust but they must still be regarded as consumable They might suffer from permanent coating of the surface and from residues formed by arcing events during fractionations Coatings of the surface may derive from substances in sample buffers that are not hold back by the gel system and can modify the electrodes Arcing during fractionation might happen if an insufficient amount of liquid is applied to pads in the assembly of the experimental setup or if these pads dry out during the fractionation Checking Fixed and Movable Electrodes 1 Make sure your two electrode sets and tray
98. s are completely dry and clean 2 For movable electrodes measure the resistance from the tip of the electrode 8 individual pins which contact to the pad to the tip of the anode connector electrode one pins 3 For movable electrodes measure the resistance from the tip of the electrode 8 individual pins which contact to the gel strip at the cathode side to the metal spring of the movable electrode 8 individual metal springs The resistance for each of this measurement should have a value below 100 kOhm In case the measured resistance is higher exchange the respective electrode See page 156 for product numbers for spare parts OFFGEL User Manual Troubleshooting 7 Troubleshooting the LAN Connection If your web interface to the Agilent 3100 OFFGEL Fractionator does not work correctly check for disconnected LAN cables at the rear of the instrument Make sure you are using the LAN connector and not the CAN connector If the problem still remains then Power down the Agilent 3100 OFFGEL Fractionator and wait 1 minute and then restart Check the positions at the rear of the instruments according to the installation instruction in chapter 3 If the problem still remains and you were had the instrument in a local network then Set up the instrument in a direct LAN connection to a PC Install a cross over LAN cable and change the configuration dip switches 7 and 8 in the rear of the instrument to On Follow the instru
99. s et les m thodes de tout bon laboratoire afin d viter la propagation de maladies L limination des solutions et produits usag s doit tre effectu e en conformit avec les directives de sant et de s curit environnementale en vigueur Manuel de l utilisateur OFFGEL 139 140 S curit Consignes de s curit Manuel de l utilisateur OFFGEL Frazionatore OFFGEL Agilent 3100 M Manuale per l utente ee e O 090 050 i Sicurezza o x B A Informazioni per la sicurezza 142 Questo capitolo contiene informazioni importanti per la sicurezza relative al sistema Frazionatore OFFGEL Agilent 3100 RE Agilent Technologies 141 Sicurezza Informazioni per la sicurezza ATTENZIONE Le seguenti norme di sicurezza generale devono essere osservate durante tutte le fasi di funzionamento manutenzione e riparazione del sistema Frazionatore OFFGEL Agilent 3100 Le istruzioni riguardanti la sicurezza devono essere lette e comprese prima dell installazione dell utilizzo e della manutenzione dello strumento Il mancato rispetto di tali norme o di avvertenze specifiche riportate nel presente manuale viola gli standard di sicurezza per progettazione produzione ed uso previsto dello strumento Agilent Technologies non puo in nessun modo essere ritenuta responsabile del mancato rispetto delle normative in vigore da parte dell utilizzatore Un segnale di allarme richiama l attenzione su una situazione che potrebbe causare d
100. s since the last instrument start It can be printed by selecting the print button or the print dialog of the browser e Posi Status Method Results Config About Time Class Number Description 09 30 46 20008 Result export succeeded 09 30 32 20008 Result export succeeded 08 04 34 109 Ready 08 03 21 20006 USB mounted 08 03 17 20007 USB not mounted 08 03 17 Cover closed 08 03 17 No pause 08 03 17 Temperature control on 08 03 17 Tray 2 focusing end 08 03 17 3 Tray 2 hold done 08 03 17 Tray 2 profile end 08 03 17 Tray 2 temperature ok 08 03 17 Tray 1 focusing end 03 03 17 Tray 1 hold done 08 03 17 Tray 1 profile end 08 03 17 Tray 1 temperature ok 08 03 17 Not ready 08 03 17 No analysis 08 03 17 Prerun Figure 16 Instrument Logbook Operation 5 The log book entries consist of date and time for an event on the instrument Each event is a ssigned a color coded class Green change in status Blue info event Red error An internal identification code serves as detailed information for the Agilent Service A brief description of the detected events is given in the last column OFFGEL User Manual 53 5 Operation Exporting Results Exporting Results The Result tab allows downloading result files from fractionation runs stored in the internal memory This can be done per individual file by using the browser download function on the results tab Also a copy of result files from a time span
101. sible Cause Action No current or very low current less than 20 of the current limit during the first 5 minutes after start of a run in one lane compared to other lanes 1 No electrical contact or impaired contact 2 IPG strip is improperly rehydrated check if there is any damage to the electrodes If so exchange the electrodes check if there is any damage to the anode connector If so call Agilent Technologies Make sure that the electrodes are in place correctly and the electrode pins are connecting to the electrode pads Make sure that the electrode pads are damp and provide a good contact between the electrode pins and the gel ends of the IPG strip Make sure that the IPG strip was rehydrated completely and that all wells in the frame are filled with rehydration buffer during rehydration One lane shows very high current signal compared to other lanes on the same tray This might impair the focusing result of all other lanes on that tray 1 the corresponding lane is leaking 2 the corresponding IPG strip was inserted in the opposite direction inspect the corresponding frame for any damage If damage is present exchange the frame Make sure that the IPG strip is inserted with the gel side up Make sure that the corresponding frame is snapped into the tray lane as described in the kit manual Make sure that the on the IPG strip is in the direction o
102. sopropanol using a lab tissue or paper towel to remove the mineral oil 5 Thoroughly air dry the anode connector or dry well with a lab tissue or paper towel prior to the next fractionation 6 Return the anode connector to its original position and fasten the screws with a hex key OFFGEL User Manual Maintenance 6 Cleaning the Top Cover and the Housing of the Instrument The top cover and the housing can be cleaned with a damp cloth either using water or a mild detergent CAUTION Do not use alcohols or ketones for the top cover since the material is not resistant to these solvents OFFGEL User Manual 75 6 Maintenance Firmware Update 76 At regular intervals Agilent Technologies releases new firmware revisions of its products to provide new features or bug fixes These revisions and the Firmware Update Tools are provided via the Agilent Technologies web page http www agilent com Life Science and Chemical Analysis Solutions Technical Support Downloads amp Utilities or direct via http www chem agilent com scripts DownloadsUtilities asp A link to this page is provided on the About page of the connected instrument see Figure 24 on page 64 This section describes how you can update the firmware of the Agilent 3100 OFFGEL Fractionator instrument It requires installation of the Firmware Update Tool installation of the firmware onto a PC connection of the PC to the instrument using a LAN cable Al
103. t Figure 8 Configuration switches of the Agilent 3100 OFFGEL Fractionator The instrument LED is usually yellow which indicates that the cooling platform is cooling down to the specified temperature typically 20 C Once the temperature has been reached the LED is switched off To test the instrument for proper functionality it is recommended to perform the tests as specified in chapter Troubleshooting the Instrument on page 92 35 4 Installation Setting up the Control PC 36 To operate the control computer delivered with the system refer to its manual You might request support from your IT department or administrator in case you do not feel comfortable with first time starting and configuring a computer Make sure to retain the administrative rights until the system setup is complete Make sure the browser Internet Explorer version 8 0 is installed The following description on necessary steps reflects a setup for computers with operating system Windows 7 Starter This description reflects steps of the Quick Installation guide G3100 90111 which is part of the instrument shipment Assignment of an IP Address to the Control Computer The Control PC and the the 3100 OFFGEL fractionator need to be enabled for LAN communication by assigning the PC a suitable IP address first This IP address is associated with a certain subnet address The 3100 OFFGEL fractionator has a similar default IP address from the same subnet
104. t Any interruption of the protective grounding will cause a potential shock hazard that could result in serious personal injury Whenever it is likely that the protection has been impaired the instrument must be made inoperative and be secured against any intended operation Any maintenance of the instrument under voltage should be avoided Do not replace components of the instruments with power turned on Do not operate the instrument in the presence of flammable gases or fumes Operation of any electrical instrument in such an environment constitutes a definite safety hazard Do not install substitute parts or make any unauthorized modification to the instrument OFFGEL User Manual 115 8 Safety Safety Symbols Table 13 Safety Symbols used on Instruments and in Manuals Symbol Description The apparatus is marked with this symbol when the user should A refer to the User Manual in order to prevent risk of harm to the operator and to protect the apparatus against damage Chemical and Biological Safety WARNING When working with solvents please observe appropriate safety procedures for example goggles safety gloves and protective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used 116 OFFGEL User Manual Safety 8 WARNING If pathogenic toxic or radioactive samples are intended to be used in this instrum
105. t or a 10 SDS solution and a brush to clean the electrodes Carefully rinse the electrodes with hand warm tap water to remove the detergent Carefully rinse the electrodes with distilled water to remove remaining salts Thoroughly air dry the electrodes prior to use to avoid any electrical short circuit Do not use heat such as a hair dryer to dry the electrodes since the plastic parts of the electrodes may be deformed CAUTION Do not use a dishwasher to clean the electrodes since the plastic parts of the electrodes may be deformed or corrosion due to alkaline conditions might occure Additional electrodes are also available as spare parts see Table 15 on page 156 Cleaning the Trays OFFGEL User Manual Trays have to be cleaned before every use to avoid cross contamination of protein or peptides from one sample to the other This is especially necessary when the instrument is used in the IPG IEF mode in gel mode 1 c A 65 N Rinse the complete tray with hand warm tap water Do not exceed a water temperature of 50 C Use a mild detergent or 10 SDS solution and a brush to clean the tray Carefully rinse the tray with hand warm tap water to remove the detergent Carefully rinse the tray with distilled water to remove remaining salts Thoroughly air dry the tray or dry well with a lint free tissue prior to use 71 6 Maintenance CAUTION Do not use heat such as a hair dryer or dish washer to clean and dry the
106. te Apply Cancel Figure 10 Tray temperature and system clock The temperature applies to both trays 42 OFFGEL User Manual Status tab Status 1336 220 C Tray I OG12PE01 v Volt Hours Time Voltage Power Current Operation 5 The Status tab shows the current state of both trays Figure 11 shows Tray lin arun The Status tab is also the main application screen which enables you to select and load methods to start and stop them The Status tab allows processing and modification of run parameters interactively of a method that is currently running Method selection Start Stop button Method Plot Logbook Results Config About Run e Tray corona Status indicator Method step Instrument lock A 20kVh 10000 hm Run parameters 4500V 200 mW 50 uA Actual values sooo ci F 65 SW so 0 aro w Actuals per lane CIS Edit Stop Figure 11 Status Screen for Tray In a pull down menu for Method Selection you can select methods that are present on the internal drive of the instrument or on the USB stick inserted at the rear of the instrument These methods can either be default methods which were supplied with the instrument or user modified methods See Chapter 5 page 65 for description of the default methods Method modification and saving can be done from the Method tab The fractionation can be started after assembly of the tray with the start button This button changes subs
107. ternatively a connection can be done by a 9 to 9 pin female connector RS 232 cable Agilent part number G1530 60600 or RS232 61601 Such a cable was included in the accessory kit see Table 4 on page 33 that was delivered with earlier systems that included a handheld controler This is a so called Null Modem Cable with full handshaking where the connection are made between pins 1 1 2 3 3 2 4 6 5 5 6 4 7 8 8 7 9 9 Besides the Firmware Update Tool you can use for upgrading future revisions of instrument Firmware also the convenient Update via USB This requires the Main Firmware B 06 30 or higher to be already installed on the instrument OFFGEL User Manual Maintenance 6 Installation of the Firmware Update Tool 1 Download the Firmware Update Tool from the Agilent web site http www chem agilent com scripts DownloadsUtilities asp onto your PC Do not use older versions of the Firmware Update Tools as provided via the Agilent web Versions below 2 3 001 introduced 07 2006 should no longer be used 2 Double click the exe file to install the program files Follow the instructions The program is installed into a folder default is C Program Files Agilent LC Firmware Update Tool You may change this according your needs A Firmware Update Tool icon is installed onto your desktop and a program folder is installed under Start Programs see Figure 27 m Agilent Firmware Update Tool PU Uninstall Firmware Upd
108. that could impair Agilent 3100 OFFGEL Fractionator functionality or performance and lead to loss of data Do not proceed beyond a caution until you have fully addressed the indicated conditions Instrument Safety This is a Safety Class instrument provided with a terminal for protective grounding and has been manufactured and tested according to international safety standards Power Cords 114 Different power cords are offered as options with the Agilent 3100 OFFGEL Fractionator The female end of all power cords is identical It plugs into the power input socket at the rear of the instrument The male end of each power cord is different and designed to match the wall socket of a particular country or region OFFGEL User Manual Safety 8 WARNING Never operate your instrument from a power outlet that has no ground connection Never use a power cord other than the Agilent Technologies power cord designed for your region WARNING Never use cables other than those supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Operation Before applying power follow the installation instructions Additionally the following must be observed Do not remove instrument covers when operating Before the instrument is switched on all protective ground terminals extension cords auto transformers and devices connected to it must be connected to a protective ground socke
109. the button Open Protocol File to open the report Information can be copied pasted into an e mail for further help from Agilent support OFFGEL User Manual 87 6 Maintenance L RES Update Tool File Action Options Help Figure 41 E FWReport txt Notepad File Edit Format View Help 4 8 2010 4 20 40 PM 4 8 2010 4 20 40 PM 4 8 2010 4 20 40 PM 4 8 2010 4 20 41 PM 4 8 2010 4 20 51 PM 4 8 2010 4 21 09 PM 4 8 2010 4 21 09 PM 4 8 2010 4 21 10 PM Select the button Open Protocol File DER Start of update Instrument 3100 OFFGEL 1 134 40 27 54 connected G3100A DE00000070 Main B 06 30 0002 Resident B 01 06 0001 Transferring G3100A DE00000070 to resident Instrument 3100 OFFGEL 1 134 40 27 54 disconnected Instrument 3100 OFFGEL 1 134 40 27 54 connected G3100A R DE00000070 Main B 06 30 0002 Resident B 01 06 0001 Update of Main firmware on G3100A R DE00000070 started B 06 30 0002 gt U 06 30 9 C Program Files Agilent LC Firmware Update Tool 3100A_U630_009 dlb 4 8 2010 4 22 44 PM 4 8 2010 4 22 44 PM 4 8 2010 4 22 52 PM 4 8 2010 4 22 57 PM 4 8 2010 4 22 57 PM Figure 42 Update of Main firmware on G3100A R DE00000070 succeeded Transferring G3100A R DE00000070 to main Instrument 3100 OFFGEL 1 134 40 27 54 disconnected Instrument 3100 OFFGEL 1 134 40 27 54 connected G3100A DE00000070 Main U 06 30 0009 Resident B 01 06 0001
110. the instrument to your local network 3 Reconfigure the instrument to use the customer set IP address by changing the configuration dip switches Figure 7 on page 34 Put switch 7 up and all other configuration dip switches down 4 Power the instrument on again The instrument will be accessible now through your local network by typing the IP address into the address line of you browser Note down the initial Network configuration of the PC for your own reference Since the instrument is not enabled for DHCP the assigned custom IP address must be a static IP address OFFGEL User Manual Agilent 3100 OFFGEL Fractionator User Manual 5 Operation The User Interface 40 General Functions 42 Status tab 43 Working with Methods 46 Displaying Data Graphically 49 Instrument Logbook 52 Exporting Results 54 Reviewing Result Files 57 Instrument Configuration 62 Additional Information 64 Performing a Fractionation 65 This chapter details how to operate the Agilent 3100 OFFGEL Fractionator It familiarizes the user with all control functions from the internet browser interface It describes the necessary steps to perform a fractionation and to change the customizable IP address of the instrument RES Agilent Technologies 39 5 Operation The User Interface 40 The Agilent 3100 OFFGEL Fractionator can be controlled from an internet browser interface The communication of browser application and the instrument is based on th
111. trays since the material may be deformed Additional trays are also available as spare parts see Table 15 on page 156 Cleaning the Cooling Platform If for any reason the cooling platform is contaminated use the following cleaning procedure 1 Wipe the cooling platform with water using a lab tissue or paper towel 2 Wipe the cooling platform with isopropanol using a lab tissue or paper towel to remove the mineral oil 3 Thoroughly air dry the cooling platform or dry well with a lab tissue or paper towel prior to the next fractionation A leak sensor is installed on the left side of the rear of the cooling platform The sensor and its surroundings has to be completely dry before it is possible to start the next fractionation Cleaning the Anode Connector If you see heavy contamination of the fixed electrode such as salts or mineral oil it is strongly recommended to also detach and clean the anode connector 72 OFFGEL User Manual OFFGEL User Manual Maintenance 6 1 Loosen the 4 screws on top of the anode connector with a hex key Figure 25 Disassembling the anode connector 2 Carefully lift the anode connector from the front Figure 26 Take care to not damage the cables at the rear of the anode connector 73 6 74 Maintenance Anode Connector Cables Figure 26 Detached anode connector 3 Wipe the anode connector with water using a lab tissue or paper towel 4 Wipe the anode connector with i
112. tre rendue impossible Eviter tout entretien de l instrument sous tension Toujours d brancher le cordon secteur pour remplacer un composant de l instrument Ne pas faire fonctionner l appareil en pr sence de gaz inflammables ni de fum e Faire fonctionner un appareil lectrique dans un tel environnement constitue un risque certain Manuel de l utilisateur OFFGEL 137 S curit Utiliser exclusivement des pi ces d origine Agilent et ne pas effectuer de modification non autoris e de l instrument Symboles de s curit Table 13 Symboles de s curit utilis s sur les instruments et dans les manuels Symbole Description L appareil est marqu de ce symbole lorsque l utilisateur doit consulter A le Manuel de l utilisateur afin de se pr munir contre les risques de blessure corporelle et de prot ger l appareil contre tout dommage S curit chimique et biologique DANGER Lors de travaux avec des solvants observez les proc dures de s curit appropriees port de lunettes gants et v tements de protection par exemple indiqu es dans la fiche technique de manipulation et de s curit du produit fournie par son fabricant tout sp cialement lorsqu il s agit de solvants toxiques et dangereux 138 Manuel de l utilisateur OFFGEL S curit DANGER Si des chantillons pathogenes toxiques ou radioactifs doivent tre utilises dans cet instrument il est de la responsabilit de l utilisateur de s assur
113. ture of the cooling platform has been reached While the main firmware is downloaded to the instrument the status indicator LED on the front of the instrument front will blink in red The status LED will blink red in case the instrument uses the resident firmware only This might happen if the main firmware installed to the instrument encountered an error or is corrupt In such case only limited functionality is available Please restart the instrument If the blinking LED is still present after an instrument restart connect to the instrument with the OFFGEL User Manual Troubleshooting 7 Firmware download Tool and update to the latest firmware revision The resident section of the firmware is identical in many Agilent devices and modules Its properties include The complete communication capabilities CAN LAN and RS 232C Memory management Ability to update the firmware of the main system Status Bars The Agilent 3100 OFFGEL interface displays several status bars see General Functions on page 42 Table9 Status bar title overview LED is yellow Status bar title Description Comment Cover open The top cover of the instrument is A running fractionation will be open paused if the top cover is opened Temp The set temperature of the cooling You can start a fractionation platform has not been reached however the status bar will switch to Wait Temp I II Wait Temp or The set temperature of the co
114. urrent in uA to Charge in mAh Modification of a currently loaded method can be done by clicking onto the Edit button or by clicking into the respective parameter area of a tray A dialog will appear and allows changing the parameter of the current method step to directly apply it to the ongoing fractionation or to cancel this attempt see Figure 12 on page 44 Upon modification an asterisk occurs next to the method name and denotes that the method was modified during the run Volt Hours kVh tine 100 00 hh mm LENA 2000 V Power mWh Current S yA Apply Cancel Figure 12 Edit a current method OFFGEL User Manual Operation 5 Status tab The instrument is accessible from any computer in the local network To protect it from unintended access an Instrument lock can be established Click onto the lock icon and select a password You might want to leave information for other users or a password hint for yourself in a free editable field The password can be set individually per tray Since it is transient it can be different from one locking event to another In case of misplaced or forgotten passwords please power cycle the instrument after the current runs are finished to release the lock Lock Tray ME Locked for Proteomics user 3 Password hint xxx Cancel Figure 13 Locking a tray OFFGEL User Manual 45 5 Operation Working with Methods Working with Methods The Method tab see Figure 14 is used to view
115. vable electrodes at the most right position 3 Load the DIAGNOSE method to both trays and press Start on both trays to start the runs 4 Check that the actual voltage values reach approximately 800 V after one minute and that the currents of all lanes are lt 1 uA If this is not the case check once again that the tray and the electrodes are completely dry and clean Inspect the cathode connector for contamination and clean it if necessary Make sure that no sparking occurs If you cannot solve this problem call Agilent Technologies for further help Otherwise proceed with step 5 5 Press Stop to stop both runs 6 Open the top cover and pipette 4 ml deionized water 18 MOhm m into lanes A C E and G on each tray 7 Close the top cover Make sure the water is evenly distributed and the electrode pins are in contact with the water However avoid spillage of water across the electrodes into neighboring lanes and the electrode connectors This might result in sparking 8 Press Start on both trays to start the DIAGNOSE method again 9 Look at both tray status screens You should see a regular current pattern as shown in Figure 43 OFFGEL User Manual 97 7 98 Troubleshooting Troubleshooting the Instrument Status Method Plot Logbook Results Config About 17 17 Run 20 0 C Tray ua d DIAGNOSE v a Volt Hours 2 5 kVh Time 0 15 h m Voltage 800 V Power 200 mw Current 50 uA mm D 5 GO
116. w chem agilent com scripts DownloadsUtilities asp onto your PC Double click the exe file to extract the content into any folder on your hard disc drive The Main Firmware file has a name starting with the instrument number revision description and the suffix dlb An example for a typical Main Firmware file name is 3100A_B630_002 dlb Create a folder on the USB stick with exactly this path X OFFGEL UPDATE Please use capital letters for the path X represents the drive letter of your USB stick and is a variable Move the extracted firmware file to the new folder on the USB stick Only exactly this file should be present in the folder Power the 3100 OFFGEL Fractionator off Insert the prepared USB stick to the USB port of the instrument Power the Agilent 3100 OFFGEL Fractionator on during the restart the instrument will check the USB stick folders for relevant files and perform an update in case the installed firmware has a lower revision Allow the instrument to perform the Firmware update A typical time for this is 10 minutes During this time the red status LED will blink After a successful update or downgrade of main or resident firmware the 3100 OFFGEL fractionator restarts automatically The instrument restart is indicated by a toggling of the three LED colors In case main and resident firmware are changed in one attempt wait until the second cycle of red blinking LED and a toggling of the three LED colors is complete
117. with conventional gels The three part diagram in Figure 1 shows starting and ending states as well as the OFFGEL fractionation process itself After rehydration of gel strips the immobilized pH gradient IPG gel seals tightly against the compartment frame Then the diluted sample is distributed across all wells in the strip Figure 1A When a voltage is applied to the ends of the gel strip the proteins or peptides move through the gel until the molecules reach a well covering a portion of the gel where the pH equals the pI of the molecule Figure 1B After fractionation the liquid fractions containing pl based separated proteins or peptides can easily be removed with a multichannel pipette and processed for downstream experiments Figure 1C OFFGEL User Manual Product overview 1 Intended Use hv A Micro wells filled with sample diluted in buffer B C IPG gel pH gradient Figure 1 Fractionation principle of the Agilent 3100 OFFGEL Fractionator OFFGEL User Manual 15 1 Product overview Features 16 pl based OFFGEL fractionation with in solution recovery for easy transfer to downstream methods such as LC MS Reproducible pI based OFFGEL fractionation down to 0 1 pH resolution depending on the consumable kit used Preconfigured methods for OFFGEL and traditional in gel IEF protein and peptide fractionation Compatible with other upstream or downstream methods immunodepletion LC MS gel based analysis

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