SE260 User Manual
Contents
1. Attach one end of each length of tubing to a cooling core 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 Install the upper buffer chamber core First steady the lower chamber with one hand and then hold the core with the other hand position it on the positioning tabs and press down listening for the core to snap into place Alternatively depress both release tabs at either side position the core on the positioning tabs press into place and release the tabs Check that the core is secure Fig 2 Core installation Wi and removal To install the core position it over the positioning tabs Then either press down listening for the core to snap into place depress both release tabs set the core in place and release To remove the core depress both release tabs and lift Release tabs 2 Coolant port 2 Fig 3a b Gel sandwich installation o Fig 3a A 10 x 8 cm gel sandwich fits flush with the bottom of the upper buffer chamber core Fig 3b A 10 x 10 5 cm gel sandwich fits against the bottom of the lower buffer chamber 3 3 Place the gel sandwich 0 Rinse away the overlay with distilled water and drain any excess water o If installing a self cast or p2. 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 instrument est con u pour l usage de laboratoire int rieur seulement Seulement les accessoire
3. Required but not included Power supply with a minimum rating 250 V 50 mA constant current or constant voltage Color coded leads 2 SE6056 HV Safety lid SE256 Upper buffer chamber core SE254B Foam gasket SE208 SE260 Lower buffer chamber SE255D Coolant port 2 Spring clamps 4 SE252 Note All electrophoresis accessories and kits are listed in the the ordering section Note Inspect glass plates for chipped edges Use only unchipped plates to prevent leaking Notched plate 3 Operating Instructions 3 1 Prepare the gel sandwich Both precast and self cast gels can be run in the SE250 units This unit accepts gels in 10 x 8 cm plates which can be cast in a Hoefer SE215 SE245 or SE275 gel caster Each unit includes notched alumina plates and rectangular glass plates If casting your own polyacrylamide gels we recommend using a notched alumina ceramic back plate because it transfers heat 40 times more rapidly than glass For applications that are not heat sensitive a notched glass plate is available Before loading gels into the electrophoresis unit the separating gel should already be completely polymerized Clean away any gel adhering to the notched alumina back plate The stacking gel if applicable can be cast in place on the electrophoresis unit Load liquid samples after the gel sandwich is installed Important Use only water or water and lt 50 ethylene glycol as
4. 00 13 0 1 SE211A 5 1 0 5 1 50 13 0 1 SE211A 5 1 5 9 1 00 5 8 1 SE211A 9 1 0 10 0 75 4 8 1 SE211A 10 75 10 1 00 4 8 1 SE211A 10 1 0 10 1 50 4 8 1 SE211A 10 1 5 12 1 00 4 75 1 SE211A 12 1 0 15 0 75 2 9 1 SE211A 15 75 15 1 00 2 9 1 SE211A 15 1 0 15 1 50 2 9 1 SE211A 15 1 5 18 1 00 2 9 1 SE211A 18 1 0 1 1 0 75 68 5 1 SE211A R 75 1 1 1 00 68 5 1 SE211A R 1 0 1 12 1 50 68 5 1 SE211A R 1 5 Microtiter spacing PPreparative reference well e p21 product quantity code number Gel Casters For 1 or 2 gels 10 x 8 10 5 Hoefer SE245 Dual Gel Caster 1 For 5 to 10 gels 10 x 8 cm SE245 Hoefer SE215 Multiple Gel Caster Complete 1 Includes 20 rectangular glass plates 10 notched alumina plates 100 sheets of wax paper space saver plate 5 filler sheets set of filler plugs and Spacer Mate Order combs and spacers separately For 2 to 4 gels 10 x 8 cm SE215 Hoefer SE275 4 Gel Caster Complete 1 Includes 10 rectangular glass plates 4 notched alumina plates 100 sheets of wax paper space saver plate 5 filler sheets set of filler plugs and Spacer Mate Order combs and spacers separately For 2 to 4 gels 10 x 10 5 cm SE275 Hoefer SE235 4 Gel Caster Complete 1 Includes 5 rectangular glass plates 4 notched alumina plates 100 sheets wax paper space saver plate 5 filler sheets set of filler plugs and Spacer Mate Order combs and spacers separately Power Supplies SE235
5. dodecyl sulfate polyacrylamide gel electrophoresis J Biol Chem 224 4406 4412 1969 e p19 Ordering Information product quantity code number Hoefer SE260 Mini gel System Mini Vertical Unit for 2 slab gels Complete Includes basic unit 10 glass plates 10 x 10 5 cm 2 alumina notched plates well locating decal Spacer Mate SE245 Dual Gel Caster 2 each 0 75 mm thick 10 well combs and 0 75 mm thick spacer sets Electrophoresis Unit Replacement Parts 1 SE260 10A 75 Foam gasket 4 5 mm x 61 cm 1 SE208 Upper buffer chamber for SE260 1 SE254B Deep lower buffer chamber for SE260 1 SE255D Lid with cables for SE260 1 SE256 Wonder Wedge plate separation tool 1 SE1514 High voltage safety lead set 1 SE6056 HV Gel Seal 0 25 oz 1 SE6070 Spring clamps for SE260 and gel casters 4 SE252 Well locating label 2 SE212 Glass and Alumina Plates 10 x 8 cm Notched alumina plates 10 SE202N 10 Rectangular glass plates 10 SE202P 10 10 x 10 5 cm Notched alumina plates 5 SE262N 5 Rectangular glass plates 5 SE262P 5 e p20 Spacers thickness mm length cm quantity code number 0 75 8 2 SE2119T 2 75 1 00 8 2 SE2119T 2 1 0 1 50 8 2 SE2119T 2 1 5 0 75 10 5 2 SE2619T 2 75 1 00 10 5 2 SE2619T 2 1 0 1 50 10 5 2 SE2619T 2 1 5 Combs wells thickness mm width mm quantity code number 5 0 75 13 0 1 SE211A 5 75 5 1
6. 8 manual Hoefer SE260 Mini vertical gel electrophoresis unit um SE260 IM Rev BO 04 12 Afro e fe r Contents Important Informialion iicet rectore Fes deg ii Waste Electrical and Electronic Equipment WEEE seee1111112 vii 1 Gel Electrophoresis Unit Function and Description 1 2 SpecifICatlOns uci eater keep edet hehe pateant 2 3 Operating Irnstr ctions s oso et erre eer at 4 4 Care and Maintenance 111111111 14 5 Troubleshooting nende da ree tata rn da vak 15 leie lut e 17 Bibliography dosi ince ehh rs nets 19 Ordering Information 20 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 heat exchanger if so equipped Do not connect the heat exchanger to a water tap or any coolant source
7. Hoefer PS300B 300V 500 mA 90 W 1 Miscellaneous PS300B Hoefer SE100 PlateMate washing and storage unit 1 SE100 TE22 Transphor Tank Unit 1 TE22 QuickFit connector male 3 8 1 e p22 QFX3 8 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 and Tris are trademarks of ICI plc Dalton Mark VII L and Sigma are trademarks of Sigma Chemical Co RBS 35 is a trademark of Pierce Chemical Co 2012 Hoefer Inc All rights reserved Printed in the USA Hoefer
8. 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 W rmeaustauscher 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 ber 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 indebolita Questo strumento disegnato per l uso di laboratorio interno solo Solo g
9. a coolant Do not use a commercial antifreeze or any alcohol based mixture 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 3 2 Prepare the unit 0 To disassemble a fully assembled unit Remove the safety lid by pressing on the handle at the top of the upper buffer chamber core while lifting the lid by the bottom edges Empty all buffer chambers and remove any gel sandwiches Then depress both release tabs and lift out the upper buffer chamber core o Rinse the instrument before each use Before using the first time disassemble the unit completely and wash with a dilute solution of a laboratory detergent and thoroughly rinse with water and distilled water e Check the gasket Periodically remove the gray silicone rubber gasket from the core Inspect for nicks and wear If the gasket appears to be intact apply a ight film of Gel seal and replace it in the groove Avoid stretching the gasket by laying it onto the groove and pressing it into place o Optional cooling Circulating pressure must not exceed 0 8 bar 12 psi above ambient pressure Do not connect the cooling core to an unregulated coolant source such as a water tap Connect the cooling core to a circulator bath such as the RCB20 PLUS Slide hose clamps 4 total onto each end of two lengths of 8 mm 5 16 vinyl or silicone tubing
10. cula 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 dano 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 pa platsen fore koppla kraften tillgangen blyen till en kraft tillgang Vander sig alla kraft tillgang kontroller av och kopplar bort kraften blyen fore flytta s kerheten locket Cirkulerar bara vatten eller 50 50 vatten ethylene glycol genom varmen exchanger i sa utrustad fall Inte kopplar varmen exchanger till en vatten kran eller nag
11. er Desliga todos controlos de estoque de poder e desconecta os chumbos de poder antes de retirar a tampa de seguranca Circulam s gua ou 50 50 glicol de gua ethylene pelo exchanger de calor se for assim eguiparam 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 gualguer org nico solvente em gualguer parte do instrumento Org nico solvente causar agress o irrepar vel unidade Nao opera com temperaturas de buffer acima do m ximo especificou especificac es t cnicas Superaquecer causar agress o irrepar vel unidade e 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 dise ado 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 Latapa de la seguridad debe estar en el lugar antes de conectar la alimentaci n lleva a una alimentaci n Apaga todos controles de alimentaci n y desconecta los plomos del poder antes de quitar la tapa de la seguridad Cir
12. f 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 Precast gels are run under the same current and voltage conditions as self cast gels It takes about one hour to run two 7 cm x 0 75 mm Laemmli gels at 40 mA 20 mA per gel constant current Check band progress after 5 minutes and again after half an hour keeping an eye on the position of the tracking dye The run is complete when the tracking dye reaches the bottom of the gel Watch the buffer level in the upper buffer chamber and if necessary replenish it before it falls below the level of the notched plate A small volume of buffer may leak past a chipped plate or nicked gasket or it may wick out through the gel Ru x After the run 0 Important Always disconnect Once the tracking dye reaches the bottom of the gel the high voltage leads from the turn off the power supply disconnect the leads and power supply before removing remove the safety lid the lid from the unit e If coolant is circulating stop the flow and disconnect the fittings or tubing Remove the core assembly with gels attached by squeezing the release tabs and lift out the core assembly o Pour out the buffer by inverting the core assembly then remove both clamps and lift away gel sandwich es from the upper buffer chamber core e Gent
13. for 1 2 minutes Use only the highest quality reagents Conduct the separation at a lower current or voltage setting Dialyze or desalt the sample Reduce the sample volume or concentration Only use freshly deionized urea Improve dissociation of subunits by heating sample in SDS sample buffer 1 2 minutes at 100 C Add more mercaptoethanol or dithiothreitol check sample treatment Only use gels that were recently prepared Check pH values of the separating and stacking gel solutions Do not back titrate buffers Sample preparation e Heat samples for no more than 1 2 minutes at 100 C Store on ice after heating Store sample on ice before it is denatured Add protease inhibitors if necessary to prevent proteolytic degradation of sample e Store samples to be frozen in aliquots to prevent repeated freezing and thawing Store at 40 C to 80 C 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 Dispose of outdated acrylamide solutions and use only the highest grade of acrylamide When preparing samples avoid using solutions with a high sodium or potassium concentration St z ages ss G i T gt a ine SE260 results Lane 1 SDS 6H high MW standard mixture Sigma Lane 2 SDS 7 Dalton Mark VII L Sigma 10 ul per lane Gel 1296 SDS PAGE Stained with Cooma
14. gung 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 far avyttras som osorterat hush llsavfall och m ste samlas in separat Var god kontakta en auktoriserad tillverkarrepresentant f r information angaende avyttring av utrustningen e pvii 1 Gel Electrophoresis Unit Function and Description The Hoefer SE260 small format vertical slab gel unit is intended for rapid electrophoresis of protein or nucleic acid samples Most samples can be run in as little as 45 minutes and only a minimal amount of sample is required The SE260 accommodates one or two 10 x 8 cm or 10 x 10 5 cm gel sandwiches The upper buffer chamber is formed when the notched side of a gel sandwich is sealed against the silicone rubber gasket The upper buffer chamber core serves as a heat exchanger if cooling is
15. heden e 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 rke Tietoa Finnish Jos tata varusteita k ytet n tavassa ei m ritetty Hoefer
16. jowym 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 nao especificada por Hoefer Inc que a protecc o fornecida pelo equipamento pode ser comprometida Este instrumento projectado para uso de interior de laborat rio s S6 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 seguranca registrada por um nacionalmente reconhecido testando laborat rio A tampa de seguranca deve estar em lugar antes de ligar o estoque de poder leva a um estoque de pod
17. lde og betjene dette produktet bruker Bare en kraftforsyning som er CE merket eller sikkerhet som ha blitt sertifisert av et som e piv nasjonalt ha blitt anerkjent prover laboratorium Sikkerheten lokket m 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 urzadzenie moze zostac obnizony Instrument ten jest przeznaczony do uzytku w laboratoriach kryty tylko Tylko akcesori w i czesci zatwierdzone lub dostarczone przez Hoefer Inc moga byc wykorzystane do eksploatacji utrzymania i obstugi tego produktu korzysta jedynie zasilacza ze jest nosz ce oznakowanie CE lub bezpiecze stwa uwierzytelnione przez uznane na poziomie kra
18. li 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 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 l acqua 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 e 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 causera 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 tilbeh r og deler godkjente eller forsynte ved Hoefer Inc kan bli brukt for drive vedlikeho
19. loading to remove particulates Dialyze or desalt the sample Adjust the solutions Check recipes gel concentrations solutions and dilutions For instance do not use Tris HCI instead of Tris e f the required pH of a solution is exceeded do not back titrate Prepare fresh buffer Dispose of older acrylamide solutions and use only stock of the highest quality Only use freshly deionized urea Adjust the voltage or current settings To increase or decrease the migration rate adjust the voltage or current by 25 50 Check gel preparation and polymerization Degas the stacking gel solution and avoid trapping air bubbles under the comb teeth Overlay the running gel with water saturated n butanol before polymerization begins to avoid forming an uneven gel surface Check sample preparation Dialyze or desalt the sample Centrifuge or filter sample before loading to remove particulates e p15 problem solution Stained sample collects Poor band resolution Bromophenol blue doesn t sharpen into a concentrated zone in the stacking gel e pl6 Near the buffer front Protein is not sufficiently restricted by the resolving gel increase the T Near the top of the gel when the buffer front has reached the bottom The gel pore size is too small Decrease the T of the resolving gel The protein has precipitated Heat the sample at a lower temperature 70 C or less
20. ly loosen and then slide away both spacers Slip an extra spacer or a Hoefer Wonder Wedge into the bottom edge to prevent breaking the ears of the notched plates and separate the plates The gel usually adheres to the alumina plate Carefully lift the gel from the plate and lay it into a tray of stain or fixative e p13 e pl4 4 Care and Maintenance Do not autoclave or heat any part above 45 C Do not use organic solvents abrasives strong cleaning solutions or strong acids or bases to clean the chambers Immediately after each use rinse the unit with water and then rinse thoroughly with distilled water Handle the upper buffer chamber core with care to prevent damage to the banana plugs Allow to air dry Clean glass and alumina 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 5 Troubleshooting problem solution Smile effect on the buffer front Protein streaks vertically Unusually slow or fast run Bands are skewed or distorted To reduce the running temperature Circulate coolant through the upper buffer chamber core Prechill the buffer Decrease the current or voltage setting 10 mA per 0 75 mm gel 15 mA per 1 5 mm thick gel Run the gel in the cold room Centrifuge or filter sample before
21. optional Attach tubing to ports on both sides of the core before attaching gel sandwiches Circulate coolant Note Stacking gel resolution is optimal when poured just before electrophoresis 3 4 Sample preparation and loading 0 If wells are already in place skip to step 2 If applicable cast the stacking gel in the unit Calculate the stacking gel monomer solution volume measure the distance in cm from the top of the resolving gel to the notch in the alumina plate This should be at least 2 cm more if the sample depth in the well is unusually high Multiply this distance by the gel width 8 3 cm and the gel thickness cm This product is the required volume in ml Deaerate the stacking gel monomer solution add catalyst and initiator and then pour Use a pipette to deliver the solution into one corner of the plate taking care not to trap any bubbles Insert a comb at a slight angle to prevent trapping air into the sandwich allowing the comb sides to rest on the spacers Overlay each gel with a thin layer of water saturated n butanol water or diluted gel buffer to prevent gel exposure to oxygen S owly 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 Allow a minimum of one hour for the gel to polymerize o Prepare the sample Increase liquid sample densit
22. ot kylmedel k lla dar 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 EXE French R German R Ea Italian R zm Spanish 14 Swedish R Waste Electrical and Electronic Eguipment WEEE This symbol indicates that the waste of electrical and electronic eguipment 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 eguipment 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 Bitte nehmen Sie Kontakt mit einem autorisierten Beauftragten des Herstellers auf um Informationen hinsichtlich der Entsor
23. p 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 lt igtig Information Danish Hvis dette udstyr bruges i en m de ikke specificeret ved Hoefer Inc den beskyttelse som er blevet forsynet af udstyret kan m ske svaekkes Dette instrument er designet for indendors laboratoriumbrug bare Bare tilbeh r og del godkendede eller forsynede ved Hoefer Inc kan m ske 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 m vaere pa plads for forbinding stramforsyningsblyet til en stremforsyning Drejer alle stramforsyningskontroller af og afbryder kraftblyet for ferning sikkerhedlaget Cirkulerer bare vand eller 50 50 vand ethylene glykol gennem varmeveksleren i s fald udrustet Forbind ikke varmeveksleren til en vandhane eller nogen kelemiddelkilde hvor vandtrykket er unregulated Introducerer Aldrig antifreeze eller noget organisk opl sningsmiddel ind i nogen del af instrumentet Organiske opl sningsmidler vil for rsage uboelig skade til enheden Driver ikke med st dpudetemperaturer over maksimummet specificerede tekniske specifications Overheding vil for rsage uboelig skade til en
24. p18 10 T 2 6 C 0 375 M 8 8 0 1 0 05 w v 0 05 viv 4 T 2 6 C 0 125 M 6 8 0 1 0 05 0 1 w v 0 05 0 1 v v 0 025 M Tris base 0 192 M glycine 8 3 0 1 Bibliography Adams L D and Gallagher S R Two Dimensional Gel Electrophoresis Using the O Farrell System Current Protocols in Molecular Biology 10 4 1 10 4 13 1992 Gallagher S R and Smith J A Electrophoretic separation of proteins Current Protocols in Molecular Biology F A Ausubel R Brent R E Kingston D D Moore J G Seidman J A Smith and K Struhl eds 10 2 1 10 2 21 1991 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 Reisfeld R A et al Acidic buffer system for resolution of cationic proteins Nature 195 281 1962 Sasse J and Gallagher S R Staining proteins in gels Current Protocols in Molecular Biology F A Ausubel R Brent R E Kingston D D Moore J G Seidman J A Smith and K Struhl eds 10 6 1 10 6 8 1991 Towbin H et al Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose procedure and some applications Proc Natl Acad Sci USA 76 4350 4353 1979 Weber K and Osborn M The reliability of molecular weight determinators by
25. recast 10 x 8 cm gel sandwich align the bottom of the plate with the bottom of the core Fig 3a The bottom of the notched plate must cover the silicone rubber gasket If installing a self cast or precast 10 x 10 5 cm gel sandwich orient the sandwich so that the notched plate faces the gasket notches at the top Set the bottom of the sandwich on the supporting ledges in the bottom of the lower chamber and center the plate so that the gasket seals both sides Fig 3b Fig 4 Securing the gel sandwich onto the upper buffer chamber core Each sandwich requires two clamps The rounded edge of the Short jaw on the clamp fits into the groove behind the gasket and the long jaw presses on the glass plate over the spacer Clamp the sandwich in place 0 Lightly press the sandwich against the gasket and secure it to the core with one spring clamp on each side Position the jaw so that the shorter rounded jaw edge fits into the core groove and the longer edge sits on the glass plate Proper positioning is important to achieve a seal and to minimize glass breakage Slide the clamps down to the stop e Repeat step 1 for the second sandwich or if running only one gel clamp a plain glass plate on the unused side of the core to prevent a possible short circuit with the unused electrode Do not fill this chamber with buffer if no gel sandwich is in place Fit short jaw of clamp into the groove Cooling is
26. required The core is hollow and equipped with ports on either side for coolant circulation Unpacking 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 2 Specifications Gel plate size 10x 8 cm Approximate gel size 8 x 7 cm Max wattage 12 W Max voltage 500 V Max amperage 500 mA Max 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 II Dimensions w xh x d 16 5 x 16 x 16 cm 6 5 x 6 3 x 6 3 in Product certifications EN 61010 1 UL 61010A 1 CSA C22 2 1010 1 CE Certified This declaration of conformity is only valid for the instrument when it is used in laboratory locations used as delivered from Hoefer Inc except for alterations described in the user manual and connected to other CE labeled instruments or products recommended or approved by Hoefer Inc Fig 1 Main components SE260 electrophoresis unit Included but not shown Glass plates Notched alumina plates Gel seal 1 4 oz Spacer Mate Well locating decal
27. s 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 exchanger 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 l instrument Les dissolvants organiques causeront des dommages irr parables l 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 l 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 Dieses Instrument wird f r den Innenlaborgebrauch nur daf r entworfen Nur Zus tze und
28. s fewer wells they are wider and require more volume to raise the level 1 mm as shown in the following table Volume of sample pL per 1 mm depth no of comb thickness mm wells 0 75 1 0 1 5 5 9 5 12 7 19 1 9 5 8 10 3 6 4 8 7 2 15 2 2 2 9 4 4 18 2 9 Note If using precast gels check that the lower gel buffer contact surface is exposed the colored plastic tape must be removed Important Do not use antifreeze or any alcohol based mixture as these will irreparably damage the core 3 5 Final assembly 0 Fill the lower buffer chamber with running buffer The SE260 holds about 250 mL Check that the lower electrode running along the bottom of the the upper buffer chamber core is completely submerged o Place the safety lid on the unit e Plug the color coded leads into the jacks of an approved power supply such as the PS300B The red lead plugs into the red output jack and the black lead plugs into the black output jack o Optional cooling Begin circulating cold water or a chilled 50 50 water ethylene glycol solution e pll Important After initial monitoring do not leave the unit unattended for more than 45 minutes before checking the progress of the the bands and the buffer level e p12 3 6 Running the Gel Gels may be run at either constant current or constant voltage A constant current setting is traditionally used with a discontinuous buffer system so that the rate o
29. ssie Blue Running conditions 20 mA one hour Appendix The following Laemmli system is slightly modified for use with the mini vertical units The Laemmli system is the most common electrophoresis protocol for SDS denatured proteins The leading ion in this discontinuous buffer system is chloride and the trailing ion is glycine Accordingly the resolving gel and the stacking gel contain Tris Cl buffers of different concentration and pH and the electrophoresis buffer contains Tris glycine All buffers contain 0 1 SDS Polyacrylamide gel composition is indicated by two different percentages 96 T total acrylamide g acryl bis x 100 100 mL 96 C crosslinker g bis x 100 g acryl bis The total percent of acrylamide T in the separating gel which can range from 5 to 20 determines the pore size Commonly the amount of crosslinker used C is 2 6 In the following example system the resolving gel composition is 10 T 2 6 C which results in a medium pore size The stacking gel composition is 4 T 2 6 C The T in the stacking gel is lower because a larger pore size is required e pl7 Final concentrations separating gel stacking gel electrophoresis buffer Acrylamide concentration Tris Cl Tris glycine pH SDS Ammonium persulfate APS TEMED To achieve any other desired final concentration adjust the acrylamide stock and water volumes TTetramethylethylenediamine e
30. 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 zamrznut nebo jak koli organick rozpou t dla do jak koli sti z tohoto n stroje Rozpustidl m z
31. y with 10 glycerol or sucrose Add a tracking dye such as phenol red or bromophenol blue For SDS protein gels use 2X treatment buffer to denature both liquid and dry samples in a test tube To liquid protein solutions add an equal volume of 2X buffer To dry protein samples add equal volumes of buffer and ddH20 to achieve the desired concentration Heat the tube in boiling water for 90 seconds then chill it in ice until ready to use Treated samples can be stored frozen for future runs Store at 40 C to 80 C O Note The amount of protein sample added to each well depends on both the sensitivity of the staining method and the distribution of protein among separate bands With Coomassie Blue it is possible to detect 1 ug in a single band with the more sensitive silver stains it is possible to detect as little as 10 ng e plo To aid in loading samples wet the well locating decal and apply it to the front of the glass plate so that the appropriate edge outlines the sample wells Note The side wells for standards of a preparative comb correspond to the outer most wells formed by the 10 well comb o Fill the sample wells and each upper buffer chamber that will be used with running buffer One upper buffer chamber holds approximately 75 mL e Underlay the sample into the wells using a fine tipped microsyringe The width of the wells depends on the number of wells per comb If the comb ha
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