Hoefer SE640 User Manual
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Hoefer SE640 User Manual

Wide-mini dual gel electrophoresis unit
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Hoefer SE640
Wide-mini Dual Gel Electrophoresis Unit
   
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SE640-IM

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Summary of Contents for Hoefer SE640

  • Page 1 Hoefer SE640 Wide-mini Dual Gel Electrophoresis Unit     SE640-IM...

  • Page 2: Table Of Contents

    Contents Important Information ........... ii Waste Electrical and Electronic Equipment (WEEE) .......vii Gel electrophoresis unit function and description ........1 Specifications............2 Unpacking and inventory ........4 Operating instructions ..........7 Care and maintenance .........23 Customer service information ........24 Troubleshooting ...........25 Appendix A: Laemmli system gels ......29 Solutions ............31 Gel recipes ............34 Appendix B: Bibliography ........36...

  • Page 3: Important Information

    Important Information – English • Rozeslat pouze voda nebo 50/50 voda/ethyleng- lykolu prostřednictvím výměník tepla je li to vybav- • If this equipment is used in a manner not speci- ena. Nemají připojení výměník tepla s vodními fied by Hoefer, Inc. the protection provided by the setřepná nebo jakékoli chladicí kapaliny zdroje, kde equipment may be impaired. tlak vody je neregulo. • This instrument is designed for indoor laboratory • Nikdy zavést prostředek proti zamrznutí nebo use only. jakákoli organická rozpouštědla do jakékoli části z • Only accessories and parts approved or supplied tohoto nástroje. Rozpustidlům způsobí nenapravi- by Hoefer, Inc. may be used for operating, main- telné poškození jednotka! taining, and servicing this product. • Nejsou provozována s pufru teplotách nad • Only use a power supply that is CE marked or maximální stanovenou technickými specifika- safety certified by a nationally recognized testing cemi. Přehřátí způsobí nenapravitelné poškození laboratory. jednotka! • The safety lid must be in place before connecting Vigtig Information – Danish the power supply leads to a power supply. • Turn all power supply controls off and disconnect • Hvis dette udstyr bruges i en måde ikke specifice- the power leads before removing the safety lid.
  • Page 4 Belangrijke Informatie – Dutch on kansallisesti tunnustettnut testaaminen labo- ratoriota. • Indien deze uitrusting in een manier wordt • Turvallisuuskansi täytyy olla paikallaan ennen gebruikt die niet door Hoefer, Inc. is gespecificeerd yhdistäminen käyttöjännitelyijyjä käyttöjännit- de bescherming die door de uitrusting is verzorgd teeseen. kan worden geschaad. • Kiertää kaikki käyttöjännitevalvonnat ja irrottaa • Dit instrument is voor binnenlaboratoriumgebruik valtalyijyt ennen poistaminen turvallisuuskantta. enkel ontworpen. • Kiertää vain vesi tai 50/50 vesi/ethyleneä glycol • Enkel onderdelen en delen keurden goed of siinä tapauksessa varustetun lämmönvaihtimen leverden door Hoefer, Inc. kan voor het bedienen läpi. Älä yhdistä lämmönvaihdinta vesinapautuk- worden gebruikt, handhavend en onderhouden seen eikä jäähdytysnestelähteeseen, missä vesi- van dit product. paine on unregulated. • gebruik Enkel een netvoeding die CE is markeerde • Pakkasneste eikä orgaaninen liuotin välineen of veiligheid die door een is gecertificeerd die osassa ei esitele Koskaan. Orgaaniset liuottimet nationaal is herkend testene laboratorium. aiheuttavat korvaamattoman vahingon yksikköön! • Het veiligheidsdeksel moet in plaats voor het • Ei käytä puskuria yllä olevia lämpötiloja enintään verbinden van de netvoeding leidt tot een määritetyillä teknisillä täsmennyksillä. Ylikuumen-...
  • Page 5 Informazioni Importanti – Italian des dommages irréparables à l’unité! • Ne pas fonctionner avec les températures de • Se quest’apparecchiatura è usata in un modo tampon au-dessus du maximum a spécifié des specificato da Hoefer, Inc. la protezione fornito spécifications techniques. La surchauffe causera dall’apparecchiatura potrebbe essere indebolita. des dommages irréparables à l’unité ! • Questo strumento è disegnato per l’uso di labora- torio interno solo. Wichtige Informationen – German • Solo gli accessori e le parti hanno approvato o • Wenn diese Ausrüstung gewissermaßen nicht hanno fornito da Hoefer, Inc. potrebbe essere angegeben durch Hoefer, Inc. verwendet wird, usato per operare, per mantenere, e per revisionare kann der durch die Ausrüstung zur Verfügung questo prodotto. gestellte Schutz verschlechtert werden. • usa Solo un alimentatore che è CE ha marcato o la • Dieses Instrument wird für den Innenlaborge- sicurezza certificato da un nazionalmente ricon- brauch nur dafür entworfen. osciuto testando il laboratorio. • Nur Zusätze und Teile genehmigten oder lieferten • Il coperchio di sicurezza deve essere nel luogo durch Hoefer, Inc. kann für das Funktionieren, das prima di collegare i piombi di alimentatore a un Aufrechterhalten, und die Wartung dieses Produk- alimentatore.
  • Page 6 • Nie działają w buforze temperatury powyżej kraftforsyningene blyene til en kraftforsyning. maksymalnego określone specyfikacje techniczne. Przegrzania spowoduje nieodwracalne szkody dla • Vender all kraftforsyningsstyring av og frakopler jednostki! kreftene blyene før fjerning sikkerheten lokket. • Sirkulerer bare vann eller 50/50 vann/ethylene Informações Importantes –   glykol gjennom oppvarmingen veksleren i så fall Portuguese utstyrer. Ikke forbind oppvarmingen veksleren til en vanntapp eller noe kjølemiddelkilde hvor • Se este equipamento é usado numa maneira não vannet trykket er unregulated. especificada por Hoefer, Inc. que a protecção forne- • Introduserer Aldri antifreeze eller noe organisk cida pelo equipamento pode ser comprometida. løsemiddel inn i noe del av instrumentet. Organi- • Este instrumento é projectado para uso de interior ske løsemiddler vil forårsake irreparabel skade på de laboratório só. enheten ! • Só acessórios e partes aprovaram ou forneceu por • Driver med buffertemperaturer over maksimum Hoefer, Inc. pode ser usada para operar, manter, e ikke spesifiserte teknisk spesifikasjoner. Å overop- servicing este produto. pheting vil forårsake irreparabel skade på enheten ! • Só usa um estoque de poder que é CE marcou ou segurança registrada por um nacionalmente recon- Wazne Informacje – Polish...
  • Page 7 • Inför aldrig kylvätska eller något organiska • Sólo utiliza una alimentación que es CE marcó o lösningsmedel in i någon del av instrumentet. la seguridad certificada por un nacionalmente Organiskt lösningsmedel ska orsaka irreparable reconocido probando el laboratorio. skada till enheten! • La tapa de la seguridad debe estar en el lugar • Använd inte med buffert temperaturer över antes de conectar la alimentación lleva a una det högsta angivna tekniska specifikationerna. alimentación. Överhettning skulle orsaka irreparabla skador på enheten! • Apaga todos controles de alimentación y desco- necta los plomos del poder antes de quitar la tapa de la seguridad. • Circula sólo agua o 50/50 glicol de agua/etileno por el intercambiador de calor si ése es el caso equiparon. No conecte el intercambiador de calor a un toque de la agua ni cualquier fuente del líquido refrigerante donde la presión del agua está libre. • Nunca introduce anticongelante ni algún solvente orgánico en cualquier parte del instrumento. Los solventes orgánicos causarán daño irreparable a la unidad! • No opera con temperaturas de búfer encima del máximo especificó especificaciones técnicas. Reca- lentar causará daño irreparable a la unidad! Viktig Information – Swedish...

  • Page 8: Waste Electrical And Electronic Equipment (Weee)

    Waste Electrical and   Electronic Equipment (WEEE) English This symbol indicates that the waste of electrical and elec- tronic 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. French Ce symbole indique que les déchets relatifs à...

  • Page 9: Gel Electrophoresis Unit Function And Description

    The focused strips are easily transferred to the second-dimension slab gel for size separation. The SE640 gel plates are 18 cm wide and 8 cm in length. Up to four gels can be run at one time if sandwiches are paired into “club sandwiches”.

  • Page 10: Specifications

    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. •...
  • Page 11 Fig 1. Main components of the color-coded leads (2) SE640 series (see Fig 4 for caster components). Included but not shown: • Gel Seal compound, ¼ oz. • Spacer-Mate spacer positioning guide safety lid • Glass plates (6) • Wonder Wedge plate separation tool •...

  • Page 12: Unpacking And Inventory

    Unpacking and inventory Unwrap all packages carefully and compare con- Note:  Before using the first tents with the packing list, making sure all items time, disassemble the unit and wash with a dilute solution of a arrived. If any part is missing, contact your local laboratory detergent and rinse sales office.
  • Page 13 Glass plates The plates are 18 cm wide and 8 cm in length Three sets of glass plates are included with each unit. Notched divider plates, ordered separately, pair two gel sandwiches to form a “club sand- wich” so that up to four gels can be run at one time.
  • Page 14 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). Spacer-Mate spacer positioning guide Aligns spacers for sandwich assembly. Combs (May be ordered separately.) Combs are avail- able in sizes that form 10, 12, 15, 20, or 28 wells, and are available in three thicknesses: 0.75, 1.0, and 1.5 mm.

  • Page 15: Operating Instructions

    Gel casting and electrophoresis procedures follow. Included are instructions for polyacryl- amide gels (used with continuous or discontinu- ous buffer systems), and gradient gels. The gels required for the SE640 must be self- cast. The Dual Gel Caster (included) holds two gel sandwiches. Prepare the gel sandwich...
  • Page 16 Construct the gel sandwich and insert   into caster both top and bottom Prepare the caster and clamps sandwich edges must be flush with the Place the spirit level into the caster center and adjust clamp guide ridges. the leveling feet. Loosen all clamp screws and make pressure space for the sandwich by sliding the pressure plates toward the screws.
  • Page 17 Club sandwich A 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 glass plates (at the outer sides a regular sandwich, except before placing the top of the sandwich) glass plate, lay the divider plate and a second set spacers...
  • Page 18 Place the laminated gasket into the casting cradle (See Fig 4) with the foam side down. Place the clamp assembly in the casting cradle, screw side facing out. Note: When turning the cams, Insert a cam into the hole on each side of the casting it is easier to keep the caster tray with the ridge (short end) pointing up.
  • Page 19 Acrylamide gels Prepare the monomer solution and pour the gel See Appendix A for SDS-PAGE recipes. Prepare the required amount of monomer solution. De-aerate and add the initiator (ammonium persulfate, APS) and catalyst (TEMED) just prior to pouring the gel. Pipet the solution into one corner of the sandwich, taking care not to introduce any air bubbles.
  • Page 20 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.
  • Page 21 Both linear and exponential gradient gels can be poured in the dual gel caster. We recommend using a Hoefer SG Series Gradient Maker. Gra- dient gels are poured with a cannula from the top of the dual gel caster (see Fig 5). A stacking gel is then poured over the gradient gel.
  • Page 22 Pour the “light” solution into the reservoir chamber (the chamber furthest from the inlet). Open the stopcock long enough to displace air between the chambers and then close. Pour the “heavy” solution into the mixing chamber and place a stirring bar into this chamber.
  • Page 23 Sample preparation and loading The sample can be loaded either while the sand- wich is in the caster or after the upper buffer Note: With Coomassie Blue it ™ chamber is attached. When loading samples while is possible to detect 1 µg of protein in a single band. With using divider plates, the samples must be loaded the more sensitive silver stains, without the upper buffer chamber in place.
  • Page 24 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 membrane proteins to 60 °C for 20 minutes. Note:  Once the sample is in the wells, take care to not jar the Store unused sample at 4 °C.
  • Page 25 Final assembly  Upper buffer chamber 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. If running only one gel: Block the second upper buffer chamber slot by Fig 6. Attaching gel sandwiches to  installing the acrylic buffer dam included with the the upper buffer chamber.
  • Page 26 Use a pipet to carefully fill each slot above the sample wells with buffer in order to minimize disturbing the samples. Then pour 100 ml of buffer into the chamber, directing the buffer stream toward the side wall. Check that no buffer is leaking around the gasket. Lower buffer chamber Place a magnetic spin bar into the lower buffer chamber and place the unit on a magnetic stirrer.
  • Page 27 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 (–).
  • Page 28 Separating the sample Electrophoresis parameters for discontinuous   polyacrylamide gels Gels may be run at either constant current or Note:  All SE600 series models use 18-cm wide plates. The constant voltage settings. A constant current set- gel thickness determines the ting is traditionally used with a discontinuous cross section (and current buffer system so that the rate of electrophoretic requirement).
  • Page 29 Time A run is complete when the tracking dye reaches the bottom of the gel. A 1.5-mm thick Laemmli SDS gel, run at 25 mA/gel without cooling, usu- ally requires 2.5 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 con- stant throughout the run.
  • Page 30 Unscrew the clamps from the sandwiches and remove. Gently loosen and then slide away both spacers. Use the Hoefer Wonder Wedge plate separator tool to separate the plates. Note: Use only flexible plastic Carefully lift the glass plate with the gel attached.

  • Page 31: Care And Maintenance

    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 Caution!  Always unplug unit damage to the banana plugs and lower electrode from electrical supply before fin.

  • Page 32: Customer Service Information

    Customer service information Technical service and repair Hoefer, Inc. offers complete technical support for all of our products. If you have any ques- tions about how to use this product, or would IMPORTANT!  Request a copy of like to arrange to repair it, please call or fax the Hoefer, Inc.

  • Page 33: Troubleshooting

    Troubleshooting problem possible cause remedy  Gel sandwich   Dirty or damaged Plates, spacers, and the gasket must be completely clean. leaks while   components Wash if necessary. casting Replace chipped plates (especially if chipped near the spacers). Check the caster gasket for cuts or cracks and replace if necessary.
  • Page 34 problem possible cause remedy  Upper buffer   Mis-aligned parts Check that the glass plates, spacers, and clamps are aligned chamber leaks and fit snugly into the upper chamber gasket. Check that both gaskets are centered and that the position- ing ridges fit inside the grooves. Dirty or damaged Check that the gasket is not damaged or pinched.
  • Page 35 problem possible cause remedy  Bands are   Incomplete gel Degas the stacking-gel solution and avoid trapping air bub- skewed or  preparation and bles under the comb teeth. distorted polymerization Irregular interface Overlay the running gel with water-saturated butanol before between stacking polymerization begins, to avoid forming an uneven gel sur- and running gels face.
  • Page 36 problem possible cause remedy  Poor band   Running Begin electrophoresis as soon as the sample is loaded to pre- resolution conditions vent 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 reagents.

  • Page 37: Appendix A: Laemmli System Gels

    Appendix A:   Laemmli system gels The Laemmli system is the most common elec- trophoresis 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.
  • Page 38 The total percent of acrylamide (% T) in the resolving gel, which can range from 5 to 20%, determines the pore size. Commonly, the amount of crosslinker used (% C) is 2.6%. In the fol- lowing example system, the resolving gel com- position is 10% T, 2.6% C, which results in a medium pore size.

  • Page 39: Solutions

    Solutions  1.  Acrylamide stock solution  (30.8% T 2.6% C Bis, 200 ml) Acrylamide (FW 71.08) 30% w/v 60 g Bis* (FW 154.2) 0.8% w/v 1.6 g Note: Filter solutions 1– 4 Deionized H to 200.0 ml through a 0.45 µm filter. Store at 4 °C away from light. IMPORTANT! ...
  • Page 40 6.  0.375 M TrisCl, 0.1% SDS, pH 8.8  (Resolving gel overlay, 100 ml) 1.5 M Tris-Cl, pH 8.8 (Soln. #2) 0.375 M 25.0 ml 10% SDS (Soln. #4) 3.5 mM 1.0 ml Deionized H to 100.0 ml —OR— Water-saturated n-butanol  Shake n-butanol and deionized H O in a separatory funnel.
  • Page 41 8.   0 .025 M Tris, 0.192 M glycine, 0.1% SDS, pH 8.3 (Electrophoresis buffer, 5.0 liters) Tris (FW 121.1) 0.025 M 15.1 g Glycine (FW 75.07) 0.192 M 72.0 g SDS (FW 288.4) 3.5 mM 5.0 g Deionized H to 5.0 liters The pH of this buffer is approximately 8.3. Do not adjust pH. Up to 20 liters can be prepared and stored for up to 2 months.

  • Page 42: Gel Recipes

    Gel recipes  The Laemmli gel recipes are for 30 ml of a single concentration solution (enough for two 1.5 mm, 18 × 8 cm gels). Tabulated are ingredients and volumes for relatively large pore gels (7.5 – 10% T range) as well as smaller pore gels (12.5 – 15% T range).
  • Page 43 For linear gradient gels, use equal volumes of low % and high % acrylamide solutions in the 2 chambers of the gradient maker. Less APS is added to extend polymerization time, and less still is added to the higher %T solution to allow polymerization to occur from the top down.

  • Page 44: Appendix B: Bibliography

    Appendix B: 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).
  • Page 45 Two-dimensional electrophoresis Adams, L. D. and Gallagher, S. R., Two-Dimensional Gel Elec- trophoresis 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.

  • Page 46: Ordering Information

    Ordering information product  quantity  code no. SE640 Dual Vertical Unit, basic. SE640 Includes: 3 sets of glass plates, 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.
  • Page 47   Gel Caster for 1 or 2 gels: Dual Gel Caster, 1– 2 gels, 18 cm wide. SE6015 Includes: 2 blank gaskets. (One included with each SE640 unit)   Clamps and Cams Replacement thumbscrews for clamps SE6003U-2 Cams, black, for clamps with cam holes SE6005L Clamp assemblies, 8 cm SE6403U  ...
  • Page 48 Combs  number  thickness  width      of wells  (mm) (mm) quantity  code number 0.75 SE511-10-.75 1.00 SE511-10-1.0 1.50 SE511-10-1.5 0.75 SE511-12-.75 1.00 SE511-12-1.0 1.50 SE511-12-1.5 0.75 SE511-15-.75 1.00 SE511-15-1.0 1.50 SE511-15-1.5 0.75 SE511-20-.75 1.00 SE511-20-1.0 1.50 SE511-20-1.5 0.75 SE511-28-.75 1.00 SE511-28-1.0 1.50 SE511-28-1.5 Comb depth 15 mm;...
  • Page 49 (mm) quantity code no. 0.75 SE6419-2-.75 1.00 SE6419-2-1.0 1.50 SE6419-2-1.5 Companion products Hoefer SE100 Plate Mate washing and storage unit SE100 Hoefer TE62 Tank Transfer Unit TE62 Hoefer TE70XP Semi-Dry Transfer Unit TE70XP Hoefer reagents for gel casting and buffers Acrylamide, MB grade 1 kg GR141-1 bis-Acrylamide, MB grade...
  • Page 50 Hoefer, Inc. 32 Scotland Blvd, Ste. 9, Bridgewater, MA02324, USA Toll Free: 1-800-813-0488 Phone:1-508-807-4665 E-mail: support@hoeferinc.com Web: www.hoeferinc.com Hoefer is a registered trademark of Hoefer, Inc. © 2023 Hoefer, Inc. — All rights reserved.

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