Hoefer SE600/SE660
Contents
1. Hurkman W J and Tanaka C K Solubilization of Plant Membrane Proteins for Analysis by Two Dimensional Gel Electrophoresis Plant Physiology 81 802 806 1986 Mets L J and Bogorad L Two dimensional polyacrylamide gel electrophoresis an improved method for ribosomal proteins Anal Biochem Jan 57 1 200 210 1974 e p31 e p32 O Farrell P H High resolution two dimensional electrophoresis of proteins J Biol Chem May 25 250 10 4007 4021 1975 Bjellgvist B et al Isoelectric focusing in immobilized pH gradients principle methodology and some applications J Biochem Biophys Methods 6 317 339 1982 Gorg A et al The current state of two dimensional electrophoresis with immobilized pH gradients Electrophoresis 9 531 546 1988 G rg A Two dimensional electrophoresis with immobilized pH gradients current state Biochem Soc Trans 21 130 132 1993 Bjellqvist B et al Micropreparative two dimensional electrophoresis allowing the separation of samples containing milligram amounts of proteins Electrophoresis 14 1375 1378 1993 Blomberg A et al Interlaboratory reproducibility of yeast protein patterns analyzed by immobilized pH gradient two dimensional gel electrophoresis Electrophoresis 16 1935 1945 1995 Ordering Information product quantity code number for 18 x 16 cm gels SE600 Dual Cooled Vertical Unit basic 1 SE600 Inclu2. Prepare the caster and clamps Place the spirit level into the caster center and adjust the leveling feet Loosen all clamp screws and make space for the sandwich by sliding the pressure plates toward the screws e Construct gel sandwiches For each sandwich choose two perfectly clean unchipped glass plates and two spacers Lay one plate on a flat surface lay the Spacer Mate alignment template onto the plate wide side at the top of the plate place a spacer along each edge and lay the second glass plate on top Fig 3 Required number of clamps Clamp size 8 cm 16 cm SE600 2 SE660 2 2 o jo o o o o o o o jo o o o ol lo o o O o o o o o o o o jo o fe 16cm 24 cm SE600 SE660 Fig 4 Club sandwich assembly Side clamps will accommodate two spacers up to 1 5 mm thick glass plates at the outer sides of the sandwich spacers notched center plate Note Do not use silicone grease or petroleum jelly to seal the sandwich These substances are difficult to remove and ultimately cause artifacts Secure the sandwich with clamps Slide one clamp at a time along the sandwich sides Finger tighten one screw on each clamp set the sandwich upright on a flat surface and loosen the screw to align the stack Take great care in aligning to ensure a seal Finger tighten all screws Remove the Spacer Mate Long sandwiches reguire two clamp assemblies on each
3. mana Hoefer SE600 SE660 Standard Dual Cooled Gel Electrophoresis Units um SE600 IM Rev C0 06 12 H O e fe r Contents Important Information io ji Waste Electrical and Electronic Equipment WEEE ee vii Gel Electrophoresis Unit Function and E deele 1 Specia ON Sinana EPEE SA iia 2 Unpacking and Inventonm eee 4 Operating Instruchons AA 7 Prepare the gel sancdhwich AAA 7 Acrylamide Gels esaau aaa aaa aaa aaa aanaacana 11 EI let ug 13 Sample Preparation and Loading 15 Final Assembly een 17 Separating the Sample 22 Care and Maintenance AAA 25 MroubleshootiN seg crscnasrin aa bak i AG AAAA 26 BiblOS el VE 30 Ordering Information 33 pi Important Information English If this equipment is used in a manner not specified by Hoefer Inc the protection provided by the equipment may be impaired This instrument is designed for indoor laboratory use only Only accessories and parts approved or supplied by Hoefer Inc may be used for operating maintaining and servicing this product Only use a power supply that is CE marked or safety certified by a nationally recognized testing laboratory The safety lid must be in place before connecting the power supply leads to a power supply Turn all power supply controls off and disconnect the power leads before removing the safety lid Circulate only water or 50 50 water ethylene glycol through the
4. Use only water or 50 50 water ethylene glycol as coolant Never use a commercial antifreeze or any alcohol based mixture or irreparable damage to the heat exchanger will result Do not connect the heat exchanger to a water tap or any other source where the water pressure is unregulated e p21 Note All SE600 series units uses 18 cm wide plates The gel thickness determines the cross section and current requirement for constant current runs The length of the plate determines the running time Table 3 Laemmli buffer system starting point guidelines Gel thickness 1 5 mm Current per gel 25 mA constant current Starting voltage 80 90 V 220 250 V Final voltage Thicker or thinner gels reguire proportionally more or less current For example a 0 75 mm gel which is half as thick as a 1 5 mm gel reguires half as much current or 12 5 mA The current must be multiplied by the number of gels For instance if two club sandwiches are installed the four gels require four times as much current The current can be increased for faster runs if active cooling is used and it can be decreased for slower overnight runs At 25 mA per gel p22 Separating the Sample Electrophoresis parameters for discontinuous polyacrylamide gels Gels may be run at either constant current or constant voltage settings A constant current mode is traditionally used with a discontinuous buffer system so
5. rke Tietoa Finnish Jos tata varusteita k ytet n tavassa ei m ritetty Hoefer Inc suojelu ehk isty varusteille saattaa olla avuton Tama valine suunnitellaan sisalaboratoriokayt lle vain Vain lis varusteet ja osat hyv ksyiv t tai toimitti Hoefer Inc oheen voi k ytt k ytt miselle valvoalle ja servicing tama tuote Vain k ytt k ytt j nnitett joka on CE merkitsi tai turvallisuus joka on todistanut aidoksi ohi joka on kansallisesti tunnustettnut testaaminen laboratoriota Turvallisuuskansi t ytyy olla paikallaan k yttoj nnitteeseen Kiert kaikki kaytt jannitevalvonnat ja irrottaa valtalyijyt ennen poistaminen turvallisuuskantta Kiert vain vesi tai 50 50 vesi ethylene glycol siin tapauksessa varustetun limm nvaihtimen l pi l yhdist l mm nvaihdinta vesinapautukseen eik j hdytysnestel hteeseen miss vesipaine on unregulated Pakkasneste eik orgaaninen liuotin v lineen osassa ei esitele Koskaan Orgaaniset liuottimet aiheuttavat korvaamattoman vahingon yksikk n Ei k yt puskuria yll olevia l mp tiloja enint n m ritetyill teknisill t smennyksill Ylikuumeneminen aiheuttaa korvaamattoman vahingon yksikk n Information Importante French Si cet quipement est utilis dans une mani re pas sp cifi par Hoefer Inc la protection fourni par l quipement pourrait tre diminu e Cet instrum
6. 2 SE6118 2 1 5 0 75 16 2 2 SE6119 2 75 1 00 16 2 2 SE6119 2 1 0 1 50 16 2 2 SE6119 2 1 5 0 75 24 2 2 SE6619 2 75 1 00 24 2 2 SE6619 2 1 0 1 50 24 2 2 SE6619 2 1 5 Companion products Hoefer SE100 Plate Mate washing and storage unit 1 SE100 QuickFit connectors female 3 8 2 QF3 8 QuickFit connectors male 3 8 2 QFX3 8 e p38 Hoefer Inc 84 October Hill Road Holliston MA 01746 Toll Free 1 800 227 4750 Phone 1 508 893 8999 Fax 1 508 893 0176 E mail support hoeferinc com Web www hoeferinc com Hoefer is a registered trademark of Hoefer Inc Coomassie is a trademark of ICI plc 2012 Hoefer Inc All rights reserved Printed in the USA Hoefer
7. Bitte nehmen Sie Kontakt mit einem autorisierten Beauftragten des Herstellers auf um Informationen hinsichtlich der Entsorgung Ihres Ger tes zu erhalten Questo simbolo indica che i rifiuti derivanti da apparecchiature elettriche ed elettroniche non devono essere smaltiti come rifiuti municipali indifferenziati e devono invece essere raccolti separatamente Per informazioni relative alle modalit di smantellamento delle apparecchiature fuori uso contattare un rappresentante autorizzato del fabbricante Este s mbolo indica que el equipo el ctrico y electr nico no debe tirarse con los desechos dom sticos y debe tratarse por separado Contacte con el representante local del fabricante para obtener m s informaci n sobre la forma de desechar el equipo Denna symbol anger att elektriska och elektroniska utrustningar inte f r avyttras som osorterat hush llsavfall och m ste samlas in separat Var god kontakta en auktoriserad tillverkarrepresentant f r information ang ende avyttring av utrustningen e pvii Gel Electrophoresis Unit Function and Description The Hoefer SE600 series vertical slab gel electrophoresis units are intended for protein and nucleic acid electrophoresis under commonly used denaturing and non denaturing conditions Up to 28 samples can be compared on a single slab gel Applications include protein separations nucleic acid fractionation and the second dimension separation of 2 D electrophoresis
8. Dieses Instrument wird f r den Innenlaborgebrauch nur daftir entworfen Nur Zus tze und Teile genehmigten oder lieferten durch Hoefer Inc kann f r das Funktionieren das Aufrechterhalten und die Wartung dieses Produktes verwendet werden Verwenden Sie nur eine Energieversorgung die CE gekennzeichnet oder durch ein national anerkanntes Probelaboratorium bescheinigte Sicherheit ist Der Sicherheitsdeckel muss im Platz vor dem AnschlieBen der Energieversorgung sein f hrt zu einer Energieversorgung Alle Energieversorgungssteuerungen abdrehen und die Macht trennen f hrt vor dem Entfernen des Sicherheitsdeckels Nur Wasser oder 50 50 Glykol des Wassers Athylens durch den Warmeaustauscher wenn so ausgestattet in Umlauf setzen Verbinden Sie den W rmeaustauscher mit einem Wasserklaps oder jeder K hlmittel Quelle nicht wo der Wasserdruck ungeregelt wird F hren Sie nie Frostschutzmittel oder jedes organische L sungsmittel in jeden Teil des Instrumentes ein Organische L sungsmittel werden nicht wiedergutzumachenden Schaden der Einheit verursachen Mit Puffertemperaturen Uber angegebenen technischen Spezifizierungen des Maximums nicht funktionieren Die berhitzung wird nicht wiedergutzumachenden Schaden der Einheit verursachen Informazioni Importanti Italian Se quest apparecchiatura usata in un modo specificato da Hoefer Inc la protezione fornito dall apparecchiatura potrebbe essere ind
9. club sandwich divider notched SE6102D 18 x 24 cm Glass plates 2 SE6602 Glass plate club sandwich divider notched SE6602D gasket SE6009 choose the appropriate spacer and plate length for your unit universal clamp SE6003U basic caster SE6015 cam e p36 Combs number thickness width of wells mm mm quantity code number 10 0 75 8 3 1 SE511 10 75 10 1 00 8 3 1 SE511 10 1 0 10 1 50 8 3 1 SE511 10 1 5 12 0 75 7 6 1 SE511 12 75 12 1 00 7 6 1 SE511 12 1 0 12 1 50 7 6 1 SE511 12 1 5 15 0 75 5 7 1 SE511 15 75 15 1 00 5 7 1 SE511 15 1 0 15 1 50 5 7 1 SE511 15 1 5 20 0 75 4 1 1 SE511 20 75 20 1 00 4 1 1 SE511 20 1 0 20 1 50 4 1 1 SE511 20 1 5 28 0 75 2 7 1 SE511 28 75 28 1 00 2 7 1 SE511 28 1 0 28 1 50 2 7 1 SE511 28 1 5 Comb depth 15 mm all others 25 mm Preparative combs These combs are 25 mm deep adjustable to 10 or 15 mm no of wells thickness width mm prep ref mm prep ref quantity code number 1 1 0 75 121 6 1 SE511 R 75 1 1 1 00 121 6 1 SE511 R 1 0 1 1 1 50 121 6 1 SE511 R 1 5 1 2 0 75 113 6 1 SE511 DR 75 1 2 1 00 113 6 1 SE511 DR 1 0 1 2 1 50 113 6 1 SE511 DR 1 5 Adjustable comb back 1 SE511 BKA Required to convert any 25 mm deep comb to 10 or 15 mm depth e p37 Spacers thickness mm length cm width cm quantity code number 1 00 16 1 2 SE6118 2 1 0 1 50 16 1
10. considerations Anal Biochem Jun 29 3 505 514 1969 Schaegger H and Von Jagow G Tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa Anal Biochem 166 368 379 1987 Weber K and Osborn M The reliability of molecular weight determinators by dodecyl sulfate polyacrylamide gel electrophoresis J Biol Chem 224 4406 4412 1969 Two dimensional electrophoresis Adams L D and Gallagher S R Two Dimensional Gel Electrophoresis Using the O Farrell System Current Protocols in Molecular Biology Ausubel F A et al eds OSC pp 10 4 1 10 4 13 1992 Anderson N G Anderson N L and Tollaksen S L Proteins of human urine I Concentration and analysis by two dimensional electrophoresis Clin Chem Jul 25 7 1199 2210 1979 Anderson Leigh and Anderson Norman G High resolution two dimensional electrophoresis of human plasma proteins Proc Natl Acad Sci USA 74 5421 5425 1977 Anderson L Two Dimensional Electrophoresis Operation of the ISO DALT System Second Edition Large Scale Biology Press 1991 Bravo R Schafer R Willecke K MacDonald Bravo H Fey S J and Celis J E More than one third of the discernible mouse polypeptides are not expressed in a Chinese hamster mouse embryo fibroblast hybrid that retains all mouse chromosomes Proc Natl Acad Sci USA Apr 79 7 2281 2285 1982
11. that the rate of electrophoretic migration remains unchanged throughout the run Under these conditions voltage increases as the run proceeds A lower current setting is recommended for higher resolution The optimal current level must be determined empirically the main factors that must be balanced include the gel concentration and migration speed and the resulting Joule heating and band distortion Table 3 lists starting point guidelines and adjustments for gel thickness number of gels and migration rate Current Current acts on the total cross section area of all the gels because the gels are connected in parallel in the electrical circuit Thus the current setting for one gel must be multiplied by the number of gels of the same cross section run simultaneously For a gel 1 5 mm thick we suggest a starting current setting of 25 mA Two 1 5 mm gels 50 mA Note Cooling may be required to control Joule heating Voltage The starting voltage for a 1 5 mm slab gel connected to a power supply set to 25 mA is usually 80 to 90 V using the SE600 with a Laemmli discontinuous buffer system for SDS gels The final voltage is typically 250 to 400 V depending on the length of the gel See Table 3 Caution After initial monitoring do not leave the unit unattended for more than 1 h before checking the progress of the bands and the buffer level Time A run is complete when the tracking dye reaches the bottom of the gel
12. then poured over the gradient gel oo O OD oo Pouring a linear gradient gel O Assemble sandwich es into the dual gel casters as described on page 8 e Set up the monomer solution flow path Run a length of clear vinyl tubing through a peristaltic pump Attach one end of the tubing to the gradient maker outlet port and the other end to a 20 cm cannula The o d of the cannula must be less than the spacer thickness Place the cannula so that it rests at the bottom of the sandwich midway between the spacers e pl3 Optional Adjust the higher percentage acrylamide solution to 15 w v sucrose or 25 v v glycerol to improve layering e pl4 Prepare the monomer solution Calculate the volume of monomer solution needed Divide the total volume in half and prepare this volume of both the higher and lower percentage acrylamide solutions o Pour the light solution into the reservoir chamber the chamber farthest from the outlet Open the stopcock between the chambers long enough to displace the air and then close Pour the heavy solution into the mixing chamber and place a stirring bar into this chamber Place the gradient maker onto a magnetic stirrer and begin stirring at a rate that mixes well but does not introduce bubbles into the solution Mix the gradient and pump the solution into the sandwich While the solution is stirring begin pumpi
13. Banana plug gold with 2 washers 1 SE6067 Glass tube with 2 grommets 1 SE6160 5 for heat exchanger lower electrode assembly Spirit level 1 SER11 Gel Seal 1 4 oz tube 1 SE6070 safety lid with cables SE6056 upper buffer chamber SE6054 heat exchanger SE6160 model reorder SE600 SE6150 SE660 SE6650 e p34 product quantity code number Gel casters For 1 or 2 gels Dual Gel Caster basic 2 gels 18 cm wide ab SE6015 Includes 2 blank gaskets for 1 or 2 gels One included with each SE600 unit For up to 4 gels Gel Caster Kit 4 gels 18 x 16 cm or 18 x 8 cm 1 SE675 Includes 8 glass plates 3 space saver plates 5 filler sheets 100 sheets of wax paper Spacer Mate alignment template and filler plugs Order combs and spacers separately For up to 10 gels Multiple Gel Caster Kit 10 gels 18 x 16 cm 1 SE615 Includes 20 glass plates space saver plate 5 filler sheets 100 sheets of wax paper Spacer Mate alignment template and filler plugs Order combs and spacers separately p35 product quantity code number Clamps and cams Clamp and Cam Kit four 16 cm clamps and 8 black cams 1 SE6003UK Replacement thumbscrews for clamps 12 SE6003U 2 Cams black for clamps with cam holes SE6005L Clamp assemblies 16 cm SE6003U Clamp assemblies 8 cm SE6403U Glass plates 18 x 16 cm Glass plates SE6102 Glass plates low fluorescence SE6102LF Glass plate
14. Cold Spring Harbor Laboratory Cold Spring Harbor NY 2001 Sasse J and Gallagher S R Staining proteins in gels Current Protocols in Molecular Biology Ausubel F A et al eds OSC 10 6 1 10 6 8 1991 SDS Polyacrylamide Gel Electrophoresis and Isoelectric Focusing Handbook 80 6013 88 Hoefer Inc 2001 Non denaturing gel systems Reisfeld R A et al Acidic buffer system for resolution of cationic proteins Nature 195 281 1962 McLellan T Electrophoresis buffers for polyacrylamide gels at various pH values Anal Biochem 126 94 1982 Hedrick J L and Smith A J Size and charge isomer separation and estimation of molecular weights of proteins by discontinuous gel electrophoresis Arch Biochem Biophys 126 155 1968 Denaturing gel systems Laemmli U K Cleavage of structural proteins during the assembly of the head of bacteriophage T Nature 227 680 685 1970 Matsudaira P T and Burgess D R SDS microslab linear gradient polyacrylamide gel electrophoresis Anal Biochem 87 386 396 1978 Schreier M H Erni B and Staehelin T Initiation of mammalian protein synthesis I Purification and characterization of seven initiation factors J Mol Biol Nov 116 4 727 753 1977 e p30 Shapiro A L and Maizel J V Jr Molecular weight estimation of polypeptides by SDS polyacrylamide gel electrophoresis further data concerning resolving power and general
15. First dimension separation of 2 D protein electrophoresis should be performed on Immobilized pH Gradient Gels The focused strips are easily transferred to the second dimension slab gel for size separation All gel plates are 18 cm wide Table 1 on page 8 lists both models and the corresponding gel plate length 16 or 24 cm Up to four gels can be run at one time if sandwiches are paired into club sandwiches The heat exchanger allows buffer temperature control in the lower chamber Specifications Gel plate size w x h SE600 18 x 16 cm SE660 18 x 24 cm Gel size SE600 14 or 16 x 16 cm 16 cm with 1 cm wide spacers 14 cm with 2 cm wide spacers Gel size SE660 14 or 16 x 24 cm 16 cm with 1 cm wide spacers 14 cm with 2 cm wide spacers Maximum watt 50 W Maximum volt 1000 V Maximum ampere 500 mA Maximum temperature 45 C Environmental Indoor use 4 40 C operating conditions Humidity up to 80 Altitude up to 2000 m Installation category II Pollution degree 2 Dimensions SE600 w x h xd 32x 29x 14cm 12 5 x 11 5 x 5 5 in SE660 32 x 37x 14 cm 12 5 x 14 5 x 5 5 in Product certifications EN61010 1 UL61010A 1 CSA C22 2 1010 1 CE Certified This declaration of conformity is valid only when the instrument is e used in laboratory locations e used as delivered from Hoefer Inc except for alterations described in the user manual and e connected to other CE label
16. In a 16 cm gel SE600 a 1 5 mm thick Laemmli SDS gel run at 25 mA gel without cooling usually requires 3 hours Longer gels require proportionally more time separations in 24 cm gels SE660 require 8 hours Electrophoresis parameters for DNA acrylamide gels DNA gels are usually run at a constant voltage setting and since buffer systems are continuous both current and voltage readings remain constant throughout the run Running conditions are expressed in units of V cm Published running conditions vary widely but voltages in the range of 1 to 3 V cm are common for overnight runs In the SE600 we have run gels at up to 12 5 V cm Cooling is required at this voltage level Record each run Keep a record of the current or voltage setting number and thickness of gels buffer system and the starting and final current or voltage readings for each run so that results can be compared Inconsistent results for the same system and settings indicate potential problems such as leaking current incorrect buffer concentrations high salt concentrations or inconsistent chemical quality Check band progress after 5 minutes and again after an hour keeping an eye on the migration rate of the tracking dye The run is complete when the tracking dye reaches the bottom of the gel Watch the buffer level and if necessary replenish it as required to keep the top electrode submerged A small volume of buffer may leak past a nicked plate or ga
17. acza e jest nosz ce oznakowanie CE lub bezpiecze stwa uwierzytelnione przez uznane na poziomie krajowym laboratorium badawcze Bezpiecze stwo lid musi by w miejsce przed pod czeniem zasilania prowadzi do zasilania Za wszystkie r d a zasilania urz dzenia steruj ce off i od czy moc prowadzi przed odbiorem bezpiecze stwa lid Kr tylko wody lub wody 50 50 ethylene glycol wymiennik ciep a poprzez je li tak wyposa one Nie nale y po czy wymiennik ciep a woda z kranu lub jakimkolwiek ch odziwo r d a je eli ci nienie wody jest nieuregulowanych Nigdy nie wprowadza rozpuszczalnika organicznego przeciw zamarzaniu lub jakichkolwiek na dowoln cz dokumentu Rozpuszczalniki organiczne spowoduje nieodwracalne szkody dla jednostki Nie dzia aj w buforze temperatury powy ej maksymalnego okre lone specyfikacje techniczne Przegrzania spowoduje nieodwracalne szkody dla jednostki Informa es Importantes Portuguese Se este equipamento usado numa maneira n o especificada por Hoefer Inc que a protec o fornecida pelo equipamento pode ser comprometida Este instrumento projectado para uso de interior de laborat rio s S acess rios e partes aprovaram ou forneceu por Hoefer Inc pode ser usada para operar manter e servicing este produto S usa um estoque de poder que CE marcou ou seguran a registrada por um nacionalmente reconhecido testando laborat
18. aksimummet specificerede tekniske specifications Overheding vil for rsage uboelig skade til enheden O pii Belangrijke Informatie Dutch Indien deze uitrusting in een manier wordt gebruikt die niet door Hoefer Inc is gespecificeerd de bescherming die door de uitrusting is verzorgd kan worden geschaad Dit instrument is voor binnenlaboratoriumgebruik enkel ontworpen Enkel onderdelen en delen keurden goed of leverden door Hoefer Inc kan voor het bedienen worden gebruikt handhavend en onderhouden van dit product gebruik Enkel een netvoeding die CE is markeerde of veiligheid die door een is gecertificeerd die nationaal is herkend testene laboratorium Het veiligheidsdeksel moet in plaats voor het verbinden van de netvoeding leidt tot een netvoeding zijn Doe alle netvoedingscontroles Uit en koppel los de machtleiding voor het verwijderen van het veiligheidsdeksel Circuleer enkel water of 50 50 water ethyleenglycol door de hitte exchanger zo ja uitrust Verbind de hitte exchanger naar een waterkraan of koelmiddelbron niet waar de waterdruk niet geregulariseerd is Stel Nooit antivriesmiddel of organische oplosmiddelen in deel van het instrument voor Organische oplosmiddelen zullen onherstelbare schade aan de eenheid veroorzaken Bedien niet met buffertemperaturen boven het maximum specificeerde technische specificaties Oververhittend zal onherstelbare schade aan de eenheid veroorzaken T
19. cast the gel and then to run it For best results take extra care to align all components when assembling sandwiches Table 1 summarizes gel casting options SE600 Both precast gels and self cast gels can be used To self cast multiple gels kits can be ordered separately the SE615 Multiple Gel Caster Kit holds up to 10 sandwiches and the SE675 Gel Caster Kit holds up to four sandwiches See the accompanying gel caster user manual for complete instructions SE660 The 24 cm gels required for the SE660 must be self cast The Dual Gel Caster included holds two gel sandwiches both top and bottom sandwich edges must be flush with the clamp guide ridges pressure bar Fig 2 Sandwich assembly Inspect glass plates for nicks Use only unchipped plates to prevent leaking Tip Use the casting cradle to hold the sandwich during alignment Remove the laminated gasket from the cradle and instead of setting the sandwich upright on a flat surface set it into the casting cradle Table1 Gel casting options model no of gel plate gels size cm available gels and casters SE600 1 4 18 x 16 Dual Gel Caster Gel Caster Kit Multiple Gel Caster Kitt Commercially available gels SE660 1 4 18 x 24 Dual Gel Caster Two accessory notched divider plates are required to run 2 extra gels SCasts up to 4 gels Casts up to 10 gels Construct the gel sandwich and insert into caster
20. des 3 sets of glass plates four 16 cm clamp assemblies 6 cams dual gel casting stand with leveling base and level buffer dam Spacer Mate alignment template and Wonder Wedge plate separation tool Order 2 combs and 2 sets of 16 cm spacers separately SE600 Dual Cooled Vertical Unit complete 1 SE600 15 1 5 Includes basic unit plus two 15 well combs and 2 spacers 1 5 mm thick for 18 x 24 cm gels SE660 Dual Cooled Vertical Unit basic 1 SE660 Includes 3 sets of glass plates four 16 cm and four 8 cm clamp assemblies 6 cams dual gel casting stand with leveling base and level buffer dam Spacer Mate alignment template and Wonder Wedge plate separation tool Order 2 combs and 2 sets of 24 cm spacers separately SE660 Dual Cooled Vertical Unit complete 1 SE660 15 1 5 Includes basic unit plus two 15 well combs and 2 spacers 1 5 mm thick Replacement parts Wonder Wedge gel plate separation tool 1 SE1514 Slotted silicone rubber gaskets for upper buffer chamber 2 SE6008B Laminated silicone rubber gaskets for casting stand 2 SE6009 Buffer dam 1 SE6032 Upper buffer chamber for SE600 SE660 1 SE6054 Lid with high voltage leads for SE600 SE660 1 SE6056 High voltage safety lead set 1 SE6056 HV Lower buffer chamber for SE600 1 SE6150 Heat exchanger for SE600 and SE660 1 SE6160 e p33 product quantity code number Lower buffer chamber for SE660 1 SE6650 Grommets for heat exchanger lower electrode assembly 4 SE6060 6
21. e cold room Centrifuge or filter sample before loading to remove particulates Overloading Load less sample Degradation Current leakage around gel Add protease inhibitor such as PMSF Check for leaks all plates and spacers must be aligned and free of grease and cracks If used the buffer dam must be secure Sample or reagent preparation If the required pH of a solution is overshot do not back titrate Discard and prepare fresh buffer Check recipes gel concentrations and buffer dilution For instance do not use Tris HCI instead of Tris for Laemmli tank buffer Decrease the salt concentration of samples Reagent quality Dispose of older acrylamide solutions and use only stock of the highest quality Use only freshly deionized urea Voltage or current settings To increase or decrease the migration rate adjust the voltage or current by 25 50 e p27 problem Bands are skewed or distorted possible cause Incomplete gel preparation and polymerization remedy Degas the stacking gel solution and avoid trapping air bubbles under the comb teeth Irregular interface between stacking and running gels Overlay the running gel with water saturated butanol before polymerization begins to avoid forming an uneven gel surface Sample preparation Stained sample collects Near the buffer front Gel concentration Degradation Dialyze or desalt the sample M
22. e proteins to 60 C for 20 minutes Store unused sample at 4 C o Underlay the sample into the wells using a fine tipped microsyringe or gel loading pipette tip Table 2 Sample volume for standard comb sizes volume of sample ul per 1 mm depth no of comb thickness mm wells 0 75 1 0 15 10 6 2 8 3 12 4 12 5 8 7 7 11 5 15 4 3 5 7 8 6 20 3 1 4 1 6 2 28 2 1 2 7 4 1 1 1 ref prep 4 90 6 121 9 183 1 2 ref prep 4 85 6 112 9 171 Fig 7 Attaching gel sandwiches to the upper buffer chamber If the assembly leaks take it to a sink and partially release the cams to allow buffer to drain out of the upper chamber Disassemble check alignment of all sandwich components and adjust if necessary A Remove cams from the lower cam holes Place the upper chamber onto the sandwiches and then insert the cams into the upper cam holes ridge short end pointing down B The final cam position not shown must be vertical so that the assembly fits into the lower buffer chamber Note Do not force the cams If you encounter unusual resistance disassemble and inspect clamp and glass alignment along the top of the sandwich Align and reinstall Final Assembly Upper buffer chamber o Rinse both buffer chambers with water and distilled water thoroughly before each use Clean away any gel adhering to the exterior of the gel sandwiches a If running only one gel Block the second up
23. e upper chamber Spacers May be ordered separately Spacers determine the thickness of the gel and are available in three thicknesses 0 75 1 0 and 1 5 mm and two widths 1 0 and 2 0 cm Spacer Mate spacer positioning guide Aligns spacers for sandwich assembly Combs May be ordered separately Combs are available in sizes that form 10 12 15 20 or 28 wells Preparative combs include 1 or 2 reference wells in addition to a preparative well Most combs are available in all three thicknesses 0 75 1 0 and 1 5 mm All preparative combs and 10 12 15 and 20 well combs form wells that are 25 mm deep The 28 well comb forms wells that are only 15 mm deep so that wells do not collapse when the comb is removed The sample volume held by each well depends on the gel thickness well depth and the number of wells per comb Table 2 lists sample volumes of each well for all combs Wonder Wedge plate separator tool Used to disassemble gel sandwiches and to gauge spacer and comb thickness Operating Instructions Gel casting and electrophoresis procedures follow Assembly of both models is identical Included are instructions for polyacrylamide gels used with continuous or discontinuous buffer systems and gradient gels See page 30 for the Bibliography Prepare the gel sandwich Glass plates spacers and clamp sets are sized so that the assembled sandwich can be easily aligned to create the seal required first to
24. ebolita Questo strumento disegnato per l uso di laboratorio interno solo Solo gli accessori e le parti hanno approvato o hanno fornito da Hoefer Inc potrebbe essere usato per operare per mantenere e per revisionare questo prodotto usa Solo un alimentatore che CE ha marcato o la sicurezza certificato da un nazionalmente riconosciuto testando il laboratorio Il coperchio di sicurezza deve essere nel luogo prima di collegare i piombi di alimentatore a un alimentatore Spegne tutto i controlli di alimentatore e disinserisce i piombi di potere prima di togliere il coperchio di sicurezza Circola solo acgua o 50 50 glicole di acqua etilene attraverso lo scambiatore di calore se cosi equipaggiato Non collegare lo scambiatore di calore a un rubinetto di acqua o qualunque fonte di refrigerante dove la pressione di acqua sregolata Non introduce mai l antigelo o qualunque solvente organico in qualunque parte dello strumento solventi organici causeranno il danno irreparabile all unit Non opera con le temperature di tampone al di sopra del massimo ha specificato le descrizioni tecniche Il surriscaldamento causer il danno irreparabile all unit Viktig Informasjon Norwegian Hvis dette utstyret blir brukt i en m te ikke spesifisert ved Hoefer Inc beskyttelsen som ha blitt git av utstyret kan bli svekket Dette instrumentet er utformet for innend rs laboratoriumbruk bare Bare
25. ed instruments or products recommended or approved by Hoefer Inc Fig 1 Main components of the SE600 series see Fig 5 for caster components Included but not shown e Gel Seal compound 1 4 oz e Spacer Mate spacer positioning guide e Glass plates 6 e Wonder Wedge plate separation tool e Buffer dam Complete unit also includes spacers 4 and combs 2 Required but not included e Magnetic stirrer e Power supply with a minimum rating of 300 V 100 mA constant A or V Optional Circulator bath Note The ordering section lists all accessories and replacement parts L color coded leads 2 safety lid upper buffer chamber with upper electrode heat exchanger with lower electrode lower buffer chamber Note Before using the first time disassemble the unit and wash with a dilute solution of a laboratory detergent and rinse thoroughly first with water and then with distilled water Unpacking and Inventory Unwrap all packages carefully and compare contents with the packing list making sure all items arrived If any part is missing contact your local sales office Inspect all components for damage that may have occurred while the unit was in transit If any part appears damaged contact the carrier immediately Be sure to keep all packing material for damage claims or to use should it become necessary to return the unit Lower buffer chamber The lower b
26. el sandwich leaks while casting Sample wells damaged or irregular Incomplete gel polymerization possible cause Dirty or damaged components remedy Plates spacers and the gasket must be completely clean Wash if necessary Replace chipped plates especially if chipped near the spacers Check the caster gasket for cuts or cracks and replace if necessary Mis aligned parts Check plate and spacer alignment realign if necessary Over clamping Air bubbles Turn cam only as far as necessary to create a seal usually 90 150 but up to 180 On each spacer apply a light film of Gel Seal compound to the bottom outside corner only Do not use silicone grease Remove air bubbles before inserting combs Slide comb into solution at an angle If comb must be removed add more monomer solution before reinserting the comb Incomplete or delayed polymerization Allow acrylamide gels to set for a minimum of 1 h Debris in wells Rinse out unpolymerized gel with sample buffer Comb removal Chemicals Remove the comb at a slight angle and very slowly to prevent damaging the gel Agarose gels Lower the comb no more than 1 cm into the gel Use only recent stocks of the highest quality reagents If the dry ammonium persulfate does not crackle when added to water replace with fresh stock Increase TEMED or APS concentration or both pH Solutions with extreme pH values especiall
27. ent est con u pour l usage de laboratoire int rieur seulement Seulement les accessoires et les parties ont approuv ou ont fourni par Hoefer Inc pourrait tre utilis pour fonctionner maintenir et entretenir ce produit utilise Seulement une alimentation qui est CET a marqu ou la s curit certifi par un nationalement reconnu essayant le laboratoire Le couvercle de s curit doit tre sa place avant connecter l alimentation mene une alimentation Tourner tous contr les d alimentation de et d brancher les avances de pouvoir avant enlever le couvercle de s curit Circuler seulement de l eau ou 50 50 glycol d eau thyl ne par l exchanger de chaleur si si quip Ne pas connecter l lexchanger de chaleur un robinet d eau ou la source d agent de refroidissement o la pression d eau est non r gul e Ne Jamais introduire d antigel ou du dissolvant organique dans n importe quelle partie de e piii instrument Les dissolvants organiques causeront des dommages irr parables a I unit Ne pas fonctionner avec les temp ratures de tampon au dessus du maximum a sp cifi des sp cifications techniques La surchauffe causera des dommages irr parables I unit Wichtige Informationen German Wenn diese Ausr stung gewisserma en nicht angegeben durch Hoefer Inc verwendet wird kann der durch die Ausr stung zur Verf gung gestellte Schutz verschlechtert werden
28. ents Poor stacking Use only gels that were recently prepared Add a stacking gel or increase height of the stacking gel Prepare the resolving gel surface by first rinsing it with stacking gel monomer before pouring the stacking gel to ensure continuity between the gels Check pH values of the resolving and stacking gel solutions Do not back titrate buffers Incomplete gel polymerization Allow gel to polymerize fully Sample preparation Store sample on ice before it is denatured Dialyze or desalt the sample Heat samples in SDS sample buffer for no more than 1 2 min at 100 C to improve dissociation of subunits Store on ice after heating Adjust the sample volume or concentration Add more mercaptoethanol or dithiothreitol check sample treatment Add protease inhibitors such as PMSF if necessary to prevent proteolytic degradation of sample Increase glycerol or sucrose to increase sample density Store samples to be frozen in aliquots to avoid repeated freeze thawing Store at 40 to 80 C e p29 Bibliography General Gallagher S R and Smith J A Electrophoretic separation of proteins In Current Protocols in Molecular Biology Ausubel F A eds OSC 10 2 1 10 2 21 1991 Hames B D and Rickwood D Gel Electrophoresis of Proteins A Practical Approach Second edition City IRL Press 1990 Sambrook J and Russell D W Molecular Cloning A Laboratory Manual
29. g care not to trap air bubbles under the teeth Club sandwich Pipette the solution into both sandwiches filling each to the same level below the notched edge Stacking gel Fill solution to 3 4 cm below the top of the glass plate This height allows 1 cm of stacking gel below the wells Pour the gel and apply an overlay see step 2 After the gel is set prepare the stacking gel as described below 2 D electrophoresis Discontinuous protein system Fill monomer solution to about 1 cm below the top of the glass plate to allow 4 5 mm for the IPG strip or tube gel and an agarose seal A stacking gel will require extra space Seal the IPG strip or tube gel in place with agarose dissolved in running buffer Take care to avoid trapping any air bubbles between the first and second dimension gels e pll Wen ij e p12 Overlay each gel with a thin layer of water saturated butanol water or diluted gel buffer to prevent gel exposure to oxygen S ow y deliver the overlay solution from a glass syringe fitted with a 22 gauge needle Apply the solution near the spacer at one side of the sandwich and allow it to flow across the surface unaided Allow the gel to polymerize for a minimum of 1 hour Stacking gel preparation Pour the stacking gel while the sandwich is still in the gel caster Stacking gel resolution is optimal when poured just before electrophoresis Remove the overlay by rinsing the top of the gel se
30. heat exchanger if so equipped Do not connect the heat exchanger to a water tap or any coolant source where the water pressure is unregulated Never introduce antifreeze or any organic solvent into any part of the instrument Organic solvents will cause irreparable damage to the unit Do not operate with buffer temperatures above the maximum specified technical specifications Overheating will cause irreparable damage to the unit Dulezit Informace Czech Pokud by toto za zen je pou ito zp sobem kter nen podle Hoefer Inc ochrana poskytovan na z klad za zen m e b t naru ena Tento n stroj je ur en pro vnit n pou it v laborato i pouze Pouze p slu enstv a sti schv len nebo poskytnut ch Hoefer Inc mohou b t pou ity pro provoz dr bu a dr b tohoto v robku zdroj nap jen pou vaj jen e je opat en ozna en m CE osv d ena nebo bezpe nost vnitrost tn uznan mi zku ebn mi laborato Bezpe nosti lid mus b t zavedena p ed p ipojen m nap jec zdroj nap jen vede k Turn ve ker nap jen kontroly vypnuto a odpojit p ed odb rem energie vede bezpe nostn v ko Rozeslat pouze voda nebo 50 50 voda ethylenglykolu prost ednictv m v m n k tepla je li to vybavena Nemaj p ipojen v m n k tepla s vodn mi set epn nebo jak koli chladic kapaliny zdroje kde tlak vody je neregulo Nikdy zav st prost edek proti zamrzn
31. in the casting stand by the upper buffer chamber and carefully lower it into the lower chamber o Inspect the installation and check the buffer levels Upper buffer chamber UBC The electrode along the upper chamber ridge must be submerged about 1 cm This level reguires 450 600 ml of buffer just enough to cover the upper chamber ribs but not high enough to contact the banana plug Lower buffer chamber LBC The lower buffer level depends on running conditions and the buffer system as described in Fig 9 Place the safety lid on the unit O Plug the color coded leads into the jacks of an approved power supply Plug the red lead into the red output jack and the black lead into the black output jack In most systems the red lead which is connected to the bottom electrode is the anode and the black lead connected to the top electrode is the cathode upper chamber buffer level lower chamber buffer level p20 Important assembly notes IEF Runs The buffer level in the lower buffer chamber must never reach the upper buffer chamber maintain at least 2 cm of clearance Do not fill the upper or lower chamber above the recommended levels illustrated in Fig 9 Remove buffer in contact with the electrode posts Pour buffer slowly and away from the slots in the upper buffer chamber to avoid disturbing the samples
32. is at the top O Note When turning the cams it is easier to keep the caster balanced if you turn both toward nsert a cam into the hole on each side of the casting tray with the ridge short end pointing up Seal the the center of the caster Fig 5 Caster components and setup gasket foam side down Y p m zzz e plo gel sandwich against the casting gasket by turning both cams as far as needed usually 90 to 150 up to 180 The cam action presses the plates down into the gasket and to seal the bottom of the sandwich The seal is complete once the glass edge appears darker and nearly transparent against the gasket Do not turn the cam past this point glass plate spacer clamp the number required depends on the plate length casting cradles leveling feet cam install ridge end up Acrylamide Gels 0 Prepare the monomer solution and pour the gel Prepare the required amount of monomer solution Deaerate and add the initiator and catalyst just prior to pouring the gel Pipette the solution into one corner of the sandwich taking care not to introduce any air bubbles See below for the appropriate solution level according to the application No stacking gel Continuous system Fill solution to just below the top of the upper plate edge If bubbles are trapped remove with a pipette or syringe Introduce a comb at a slight angle into each sandwich takin
33. jacks in the power supply Always install the safety lid before use Glass plates All plates are 18 cm wide but each model accommodates a specific plate length The SE600 takes 16 cm plates and the SE660 takes 24 cm plates Three sets of glass plates are included with each unit Notched divider plates ordered separately pair two gel sandwiches to form a club sandwich so that up to four gels can be run at one time Clamps Two 16 cm clamps are used to secure any length sandwich An additional pair of 8 cm clamps are required to secure 24 cm sandwiches The clamp pressure bar adjusted with screws distributes pressure evenly Casting stand The casting stand holds assembled gel sandwiches upright for casting gels Adjustable feet level the caster laminated gasket in the bottom of each casting cradle seals the bottom of the sandwich when it is cammed into the stand Cams Cams are used twice first to secure the assembled sandwich in the casting stand and second to attach the sandwich to the upper buffer chamber ep Rubber gaskets There are two sets of two gaskets The solid laminated gaskets fit into the bottom of the casting stand and form the seal for casting the gel The slotted gaskets fit under the upper buffer chamber and form the seal between the upper and lower chambers The ridges on the upper gasket align the gasket slot to maintain an open channel between the top of the gel and the buffer in th
34. lvents abrasives strong cleaning solutions or strong acids or bases to clean the chambers e Do not soak the laminated gasket Note If the old tube is cracked or broken protect your hand with thick gloves a piece of cloth or paper towel before removing the tube Care and Maintenance Cleaning Immediately after each use rinse the upper and lower buffer chambers with water and then rinse thoroughly with distilled water Handle the upper buffer chamber with care to prevent damaging the banana plug Clean gaskets with mild detergent and rinse with distilled water Allow to air dry Clean glass plates and spacers with a dilute solution of a laboratory cleanser such as RBS 35 then rinse thoroughly with tap and distilled water Glass plates can also be treated with but not stored in acid cleaning solutions Replacing a heat exchanger glass tube o Remove the tube by simultaneously twisting and sliding it down as far as possible until the top end is free of the upper grommet Carefully guide the tube so that it will clear the assembly then lift the tube out of the lower grommet e Lightly grease the outside of both ends of the new tube with silicone grease Twist and slide one end of the tube into the lower grommet Then slip the other end into the top grommet gently pushing it with a slight twist until it stops Check that the grommet is not pinched e p25 Troubleshooting problem G
35. n temperature by the amount indicated on the graph This should be checked at three points 0 1 v 2 E 3 o 8 a 5 3 4 lt 5 5 o 6 7 0 10 20 30 40 50 60 power supply setting W Example Run parameters 200 V 0 05 A 50 mA 1 Calculate W if your power supply does not display power directly W VxA 10 W 200 V x 0 05 A 2 Interpolate the number of degrees to subtract from the desired run temperature 10 W intersects the graph at about 1 C If the desired temperature is 23 C set the bath to 23 1 22 C If the desired temperature is 4 C set the bath to 4 1 3 C e pl9 EE Fig 9 Buffer chamber levels Approximate volumes required SE600 4 5 liters SE660 7 0 liters e Low voltage runs e Systems using the same buffer in both the upper and lower chambers upper and lower chamber buffer levels are egual Approximate volumes reguired SE600 4 0 liters SE660 6 0 liters e High voltage runs e Isoelectric focusing e Systems using different buffers in the upper and lower chambers The buffer level should cover most of the gel sandwich to allow the dissipation of heat but must allow at least 2 cm clearance below the upper buffer chamber Fit the upper buffer chamber assembly into the lower buffer chamber Use a steady hand to avoid disturbing the samples Grasp the assembly
36. ng from the mixing chamber and open the stopcock to the reservoir chamber Raise the cannula as liguid enters the sandwich keeping the tip at the gel surface Prepare more gels as reguired O Overlay each gel with a thin layer of water saturated n butanol water or diluted gel buffer to prevent gel exposure to oxygen Slowly deliver the overlay solution from a glass syringe fitted with a 22 gauge needle Apply the solution near the spacer at the side of the sandwich and allow it to flow across the surface unaided o Allow the gels to polymerize for a minimum of 1 h After polymerization pour off the overlay and rinse the gel surface several times with distilled water Prepare the stacking gel monomer solution pour the stacking gel and introduce a comb at a slight angle into the sandwich taking care not to trap air under the teeth Allow a minimum of 1 h for the gel to polymerize Note With Coomassie Blue it is possible to detect 1 ug of protein in a single band With the more sensitive silver stains it is possible to detect as little as 10 ng of protein Sample Preparation and Loading The sample can be loaded either while the sandwich is in the caster or after the upper buffer chamber is attached When loading samples while using divider plates the samples must be loaded without the upper buffer chamber in place The amount of sample loaded depends on the thickness of the gel the sensitivity
37. of the detection method used and the amount of sample expected in each band In a continuous buffer system the protein sample should be relatively concentrated because no stacking gel is used In a discontinuous buffer system the zone into which each molecular species migrates is sharpened by the stacking gel so the sample need not be as concentrated O Prepare the wells Remove the comb by gently rocking it side to side and then lifting it straight up to avoid damaging the well walls Carefully rinse each well with distilled water to remove unpolymerized acrylamide and then drain by inverting the gel sandwich or caster Fill each well with electrophoresis buffer a Prepare the sample Increase liguid sample density with 10 glycerol or sucrose Add a tracking dye such as phenol red bromophenol blue or pyronin Y For SDS protein gels use 2X treatment buffer to denature both liguid and dry samples in a test tube To liguid protein solutions add an egual volume of 2X buffer To dry protein samples add egual volumes of 2X sample buffer and high purity water to achieve the desired concentration e p15 Note Once the samples are in the wells take care to not jar the sandwiches so that the samples are not spilled or mixed e p16 Heat the tube in boiling water for 90 seconds then allow to cool to room temperature Treated samples can be stored at 40 to 80 C for future runs Heat membran
38. olecules are not sufficiently restricted by the resolving gel pore size increase the T Proteins may be degraded by endogenous proteases use protease inhibitors during the isolation step Near the top of the gel when the buffer front has reached the bottom Gel concentration Precipitation The gel pore size is too small decrease the T of the resolving or stacking gel The protein has precipitated Heat the sample at a lower temperature 70 C or less for 1 2 min At both top and bottom of the gel Tracking dye doesn t sharpen into a concentrated zone in the stacking gel e p28 Gel concentration Poor stacking The molecular weight range of the sample requires an acrylamide concentration gradient to resolve the full range of protein sizes Pour a taller stacking gel For best results allow a stacking gel height of 2 5 times the height of the sample in the well Reagent quality Dispose of outdated acrylamide solutions and use only the highest grade of acrylamide Sample preparation When preparing samples avoid using solutions with high salt concentrations problem possible cause Poor band Running resolution conditions remedy Begin electrophoresis as soon as the sample is loaded to prevent low molecular weight species from diffusing Conduct the separation at a lower current or voltage setting to reduce Joule heating Reagent quality Use only the highest quality reag
39. per buffer chamber slot by installing the acrylic buffer dam included with the unit Fit clamps onto the dam taking care to align the clamp ends and dam edges Install the dummy gel screws facing out in the second cradle in the dual gel caster Attach the gel sandwich to the upper buffer chamber Turn the upper buffer chamber upside down and place a slotted gasket into both sandwich holder recesses Both the slot in the gasket and the slot in the recess must align Both slotted gaskets must be used even if running only one gel sandwich Grooves along each slot help keep the gasket in place Additionally a small amount of Gel Seal can be applied at each end of the gasket before install to help hold the gasket against the upper buffer chamber Release the sandwiches from the caster by removing all bottom cams if present Lower the upper buffer chamber onto the gel sandwiches in the casting stand Install the cams ridge pointing down into the buffer chamber cam holes Clamp the sandwich in place by simultaneously turning one cam clockwise and the other counterclockwise a full 180 e p17 Ween 0 Note If the cooling option is used frequently it is convenient to attach QuickFit connectors to the tubing The valves in these fittings prevent coolant spillage e pl8 Use a pipette to carefully fill each slot above the sample wells with buffer to minimize disturbing the samples Then pour 100 ml of buffer in
40. rio A tampa de seguran a deve estar em lugar antes de ligar o estoque de poder leva a um estoque de poder Desliga todos controlos de estoque de poder e desconecta os chumbos de poder antes de retirar a tampa de seguran a Circulam s gua ou 50 50 glicol de gua ethylene pelo exchanger de calor se for assim equiparam N o ligue o exchanger de calor a uma torneira de gua nem qualquer fonte de refrigerante onde a press o de gua n o regulado Nunca introduz anticongelante nem qualquer org nico solvente em qualquer parte do instrumento Org nico solvente causar agress o irrepar vel unidade N o opera com temperaturas de buffer acima do m ximo especificou especifica es t cnicas Superaquecer causar agress o irrepar vel unidade pv Informaci n Importante Spanish Si este equipo es utilizado en una manera no especificado por Hoefer Inc la protecci n proporcionado por el equipo puede ser da ada Este instrumento es disenado para el uso interior del laboratorio s lo S lo accesorios y partes aprobaron o suministraron por Hoefer Inc puede ser utilizado para operar para mantener y para atender a este producto S lo utiliza una alimentaci n que es CE marc o la seguridad certificada por un nacionalmente reconocido probando el laboratorio La tapa de la seguridad debe estar en el lugar antes de conectar la alimentaci n lleva a una alimentaci n Apaga todo
41. s controles de alimentaci n y desconecta los plomos del poder antes de quitar la tapa de la seguridad Circula s lo agua o 50 50 glicol de agua etileno por el intercambiador de calor si se es el caso equiparon No conecte el intercambiador de calor a un toque de la agua ni cualquier fuente del l quido refrigerante donde la presi n del agua est libre Nunca introduce anticongelante ni alg n solvente org nico en cualquier parte del instrumento Los solventes org nicos causar n da o irreparable a la unidad No opera con temperaturas de b fer encima del m ximo especific especificaciones t cnicas Recalentar causar da o irreparable a la unidad Viktig Information Swedish om denna utrustning anv nds i ett s tt som inte har specificeras av Hoefer Inc skyddet tillhandah ll vid utrustningen kan skadas Detta instrument formges f r inomhuslaboratorium anv ndning bara Bara medhj lpare och delar godk nde eller levererade vid Hoefer Inc kan anv ndas f r fungera underh lla och servicing denna produkt anv nder bara en kraft tillg ng som r CE markerade eller s kerhet intygade vid en nationellt erk nd testande laboratorium S kerheten locket m ste vara p platsen f re koppla kraften tillg ngen blyen till en kraft tillg ng V nder sig alla kraft tillg ng kontroller av och kopplar bort kraften blyen f re flytta s kerheten locket Cirkulerar bara vatten eller 50 50 vatten e
42. side When assembling 24 cm sandwiches align each end separately That is align one end finger tighten the screws turn the sandwich 180 and align the other end In each case allow the clamp to slide down and align perfectly with the top or bottom edge of the glass plates Club sandwich A 16 or 24 cm long notched center divider plate ordered separately pairs two sandwiches to double the number of gels that can be cast and run Assemble a club sandwich in the same manner as a regular sandwich except before placing the top glass plate lay the divider plate and a second set of spacers on the stack Place the notch so that it will be at the top of the gels It is essential that the spacers and plates align perfectly in order to create a seal For 24 cm long plates position the 8 cm clamp along the side at the bottom of the sandwich farthest from the notch o Remove the sandwich and inspect the bottom to make sure that edges are aligned flush in order to ensure a complete seal Adjust if necessary Optional Apply a light film of Gel Seal compound only on the bottom corner surfaces created by the spacers and plates if your sandwiches continue to leak after several attempts of alignment Ween y Place the laminated gasket into the casting cradle See Fig 5 with the foam side down Place the clamp assembly in the casting cradle screw side facing out For 24 cm plates place the sandwich so that the onger clamp
43. sket or buffer may pass through the gel e p23 Note Use only flexible plastic prying tools to avoid chipping the glass plates p24 After electrophoresis Once the tracking dye reaches the bottom of the gel turn off the power supply disconnect the leads and remove the safety lid using finger leverage between the lid and the top of the heat exchanger Lift straight up to avoid bending the banana plugs a If coolant is circulating stop the flow and disconnect the fittings or tubing Pull out the upper buffer chamber assembly Pour the buffer into a sink Install the assembly in the dual gel caster and then release the sandwiches by turning and removing the cams o Unscrew the clamps from the sandwiches and remove Gently loosen and then slide away both spacers Use the Hoefer Wonder Wedge Gel Plate Separation tool to separate the plates Carefully lift the glass plate with the gel attached Handle the gel with care to avoid damaging it Invert the plate and position the gel low over the staining tray Pry one corner of the gel away from the glass and allow it to drop into the tray or if the gel is thick enough to handle lift it and place it into the tray To avoid splashing add staining or fixative solution to the tray after the gel is transferred O Clean the unit as described in the next section e Do not autoclave or heat any part above 45 C e Do not use organic so
44. thylene glycol genom v rmen exchanger i s utrustad fall Inte kopplar v rmen exchanger till en vatten kran eller n got kylmedel k lla d r vattnet trycket r unregulated Inf r aldrig kylv tska eller n got organiska l sningsmedel in i n gon del av instrumentet Organiskt l sningsmedel ska orsaka irreparable skada till enheten Anv nd inte med buffert temperaturer ver det h gsta angivna tekniska specifikationerna verhettning skulle orsaka irreparabla skador p enheten e pvi English EEE French R German R Ea Italian R lt Spanish D Swedish R Es Waste Electrical and Electronic Equipment WEEE This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of your equipment Ce symbole indique que les d chets relatifs l quipement lectrique et lectronique ne doivent pas tre jet s comme les ordures m nag res non tri es et doivent tre collect s s par ment Contactez un repr sentant agr du fabricant pour obtenir des informations sur la mise au rebut de votre quipement Dieses Symbol kennzeichnet elektrische und elektronische Ger te die nicht mit dem gew hnlichen unsortierten Hausm ll entsorgt werden d rfen sondern separat behandelt werden m ssen
45. tilbeh r og deler godkjente eller forsynte ved Hoefer Inc kan bli brukt for drive vedlikeholde og betjene dette produktet bruker Bare en kraftforsyning som er CE merket eller sikkerhet som ha blitt sertifisert av et som O piv nasjonalt ha blitt anerkjent prover laboratorium Sikkerheten lokket ma veere pa plass for forbinding kraftforsyningene blyene til en kraftforsyning Vender all kraftforsyningsstyring av og frakopler kreftene blyene for fjerning sikkerheten lokket Sirkulerer bare vann eller 50 50 vann ethylene glykol gjennom oppvarmingen veksleren i s fall utstyrer Ikke forbind oppvarmingen veksleren til en vanntapp eller noe kjalemiddelkilde hvor vannet trykket er unregulated Introduserer Aldri antifreeze eller noe organisk losemiddel inn i noe del av instrumentet Organiske losemiddler vil for rsake irreparabel skade pa enheten Driver med buffertemperaturer over maksimum ikke spesifiserte teknisk spesifikasjoner A overoppheting vil for rsake irreparabel skade pa enheten Wazne Informacje Polish Jezeli ten sprzet jest wykorzystywany w spos b nie okreslone przez Hoefer Inc do ochrony przewidzianej przez urz dzenie moze zosta obni ony Instrument ten jest przeznaczony do u ytku w laboratoriach kryty tylko Tylko akcesori w i cz ci zatwierdzone lub dostarczone przez Hoefer Inc mog by wykorzystane do eksploatacji utrzymania i obs ugi tego produktu korzysta jedynie zasil
46. to the chamber directing the buffer stream toward the side wall Check that no buffer leaks around the gasket Lower buffer chamber Place a magnetic spin bar into the lower buffer chamber LBC and place the unit on a magnetic stirrer Fill the lower chamber with up to 4 liters of buffer a Lower the heat exchanger into the lower chamber fitting the ports into the notches in the rim The heat exchanger must be in place for all runs because the lower electrode is integrated into the heat exchanger If no cooling is reguired skip to step 3 Optional Connect the heat exchanger to a thermostatic circulator Slide hose clamps four total onto each end of two lengths of 10 12 mm i d 3 8 1 2 vinyl or silicone tubing Attach one end of each length of tubing to a heat exchanger port Attach the free ends of each length of tubing to the circulator bath ports one to the inlet and the other to the outlet Secure the connections with the hose clamps Use the chart Fig 8 on page 19 to estimate a starting point for the circulator bath temperature setting Adjust as necessary for variables such as ambient temperature changes in power output and circulator bath efficiency If accurate temperature control is critical measure the temperature and adjust as necessary Optional Prechill the buffer Fig 8 Approximate circulator bath temperature setting Set the circulator bath temperature setting lower than the desired ru
47. uffer chamber is transparent acrylic which allows visual tracking of electrophoresis progress The chamber is chemically resistant to common electrophoretic buffers but not to organic solvents or strong acids and alkali Temperatures above 45 C may cause the chamber to warp Upper buffer chamber The upper buffer chamber is molded polysulfone which is chemically resistant to common electrophoretic buffers but not to organic solvents or strong acids and alkali The upper electrode cathode runs along the center ridge and terminates at the banana plug The upper chamber requires 0 5 0 8 liters of buffer fill no higher than the top of the plastic ribs Heat exchanger The heat exchanger must be installed for every use because it houses the bottom electrode anode which runs along the bottom of the frame When connected to a circulator bath the heat exchanger regulates the buffer temperature in the lower chamber Coolant passes through the glass tubes which are secured with silicone rubber grommets The heat exchanger connector ports are 13 mm o d The heat exchanger is rated to a maximum of 0 8 atmospheres above ambient 12 psig Connect only to coolant sources with regulated pressure Do not connect to the water tap Safety lid The banana plug on the heat exchanger connects to the red lead and the plug on the upper buffer chamber connects to the black lead The 4 mm shrouded color coded leads plug into color coded
48. ut nebo jak koli organick rozpou t dla do jak koli sti z tohoto n stroje Rozpustidl m zp sob nenapraviteln po kozen jednotka Nejsou provozov na s pufru teplot ch nad maxim ln stanovenou technick mi specifikacemi P eh t zp sob nenapraviteln po kozen jednotka Z igtig Information Danish Hvis dette udstyr bruges i en made ikke specificeret ved Hoefer Inc den beskyttelse som er blevet forsynet af udstyret kan maske svaekkes Dette instrument er designet for indend rs laboratoriumbrug bare Bare tilbehor og del godkendede eller forsynede ved Hoefer Inc kan maske bruges for drive funktionsfejl og betjening dette produkt bruger Bare en stromforsyning der er CE markerede eller sikkerhed som er blevet attesteret af en som nationalt er blevet anerkendt prove laboratorium Sikkerhedlaget ma veere pa plads for forbinding stramforsyningsblyet til en str mforsyning Drejer alle stramforsyningskontroller af og afbryder kraftblyet for fjerning sikkerhedl get Cirkulerer bare vand eller 50 50 vand ethylene glykol gennem varmeveksleren i s fald udrustet Forbind ikke varmeveksleren til en vandhane eller nogen kolemiddelkilde hvor vandtrykket er unregulated Introducerer Aldrig antifreeze eller noget organisk oplosningsmiddel ind i nogen del af instrumentet Organiske opl sningsmidler vil for rsage uboelig skade til enheden Driver ikke med st dpudetemperaturer over m
49. veral times with distilled water Invert the caster to drain To ensure a seamless contact between the resolving and stacking gels remove residual liguid by blotting one corner with a lab wipe a Calculate the stacking gel monomer solution volume Prepare the stacking gel monomer solution deaerate it and add catalyst and initiator Pour the stacking gel onto the resolving gel with a disposable or Pasteur pipette to a level about 2 mm from the top of the plate o Introduce a comb at a slight angle into the sandwich taking care not to trap air under the teeth Allow a minimum of 1 hour for the gel to polymerize Fig 6 Pouring a gradient gel A pipette tip may be used instead of a cannula if the gel solution is delivered at a rate that maintains a continuous stream on the glass surface Note Gradient gels poured in the SE615 or SE675 Multiple Gel Caster are introduced through the bottom Note When pouring an exponential gradient gel position a plunger or sealing plug above the liquid in the mixing chamber to hold the volume constant Gradient Gels Both linear and exponential gradient gels can be poured in the dual gel caster We recommend using a Hoefer SG Series Gradient Maker Gradient gels are poured from the top of the caster with a cannula if using the provided dual gel caster or from the bottom if using a Hoefer Multiple Gel Caster see instructions accompanying the caster A stacking gel is
50. y acidic may not polymerize Oxygen Remove oxygen from the gel environment Degas the monomer solution 5 10 min before pouring and then overlay the gel surface with water saturated n butanol Temperature Adjust the gel solution temperature to a minimum of 20 C especially for low T gels p26 problem Upper buffer chamber leaks Power supply detects current leak Dye front curves up smiles at edges Protein streaks vertically Unusually slow or fast run possible cause Mis aligned parts remedy Check that the glass plates spacers and clamps are aligned and fit snugly into the upper chamber gasket Check that both gaskets are centered and that the positioning ridges fit inside the grooves Dirty or damaged components Electrical path to outside ground earth Uneven heat distribution Check that the gasket is not damaged or pinched Replace if necessary Check that the upper buffer chamber is not warped from prior exposure to excessive heat Add more silicone grease to seal heat exchanger grommets Check for leaks or cracks in the heat exchanger Replace worn grommets Fill the lower buffer chamber to the level appropriate for at edges the run See Fig 9 page 20 Use magnetic stirrer and stir bar to keep buffer well mixed Excessive heat Particulates in sample Circulate ext coolant Decrease the current or voltage setting Prechill the buffer Run the gel in th
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