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Summary of Contents for Hoefer SE600 Chroma
- Page 1 Hoefer SE600 Chroma Standard Dual Cooled Gel Electrophoresis Unit SE600X-IM...
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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 ....... 8 Prepare the Gel Sandwich......8 Acrylamide Gels ........12 Gradient Gels ........ -
Page 3: Important Information
Important Information – English před odběrem energie vede bezpečnostní víko. • Rozeslat pouze voda nebo 50/50 voda/ • If this equipment is used in a manner not specified ethylenglykolu prostřednictvím výměník tepla je by Hoefer, Inc. the protection provided by the li to vybavena. Nemají připojení výměník tepla s equipment may be impaired. vodními setřepná nebo jakékoli chladicí kapaliny • This instrument is designed for indoor laboratory zdroje, kde tlak vody je neregulo. use only. • Nikdy zavést prostředek proti zamrznutí nebo • Only accessories and parts approved or supplied jakákoli organická rozpouštědla do jakékoli by Hoefer, Inc. may be used for operating, části z tohoto nástroje. Rozpustidlům způsobí maintaining, and servicing this product. nenapravitelné poškození jednotka! • Only use a power supply that is CE marked or • Nejsou provozována s pufru teplotách nad safety certified by a nationally recognized testing maximální stanovenou technickými specifikacemi. laboratory. Přehřátí způsobí nenapravitelné poškození jednotka! • The safety lid must be in place before connecting the power supply leads to a power supply. Vigtig Information – Danish • Turn all power supply controls off and disconnect the power leads before removing the safety lid. - Page 4 Belangrijke Informatie – Dutch joka on kansallisesti tunnustettnut testaaminen laboratoriota. • Indien deze uitrusting in een manier wordt • Turvallisuuskansi täytyy olla paikallaan gebruikt die niet door Hoefer, Inc. is gespecificeerd ennen yhdistäminen käyttöjännitelyijyjä de bescherming die door de uitrusting is verzorgd käyttöjännitteeseen. 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 worden gebruikt, handhavend en onderhouden vesinapautukseen eikä jäähdytysnestelähteeseen, van dit product. missä vesipaine 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 verbinden van de netvoeding leidt tot een enintään määritetyillä teknisillä täsmennyksillä.
- Page 5 Informazioni Importanti – Italian l’instrument. Les dissolvants organiques causeront des dommages irréparables à l’unité! • Se quest’apparecchiatura è usata in un modo • Ne pas fonctionner avec les températures de specificato da Hoefer, Inc. la protezione fornito tampon au-dessus du maximum a spécifié des dall’apparecchiatura potrebbe essere indebolita. spécifications techniques. La surchauffe causera • Questo strumento è disegnato per l’uso di des dommages irréparables à l’unité ! laboratorio interno solo. Wichtige Informationen – German • Solo gli accessori e le parti hanno approvato o hanno fornito da Hoefer, Inc. potrebbe essere • Wenn diese Ausrüstung gewissermaßen nicht usato per operare, per mantenere, e per revisionare angegeben durch Hoefer, Inc. verwendet wird, questo prodotto. kann der durch die Ausrüstung zur Verfügung • usa Solo un alimentatore che è CE ha marcato gestellte Schutz verschlechtert werden. o la sicurezza certificato da un nazionalmente • Dieses Instrument wird für den riconosciuto testando il laboratorio. Innenlaborgebrauch nur dafür entworfen. • Il coperchio di sicurezza deve essere nel luogo • Nur Zusätze und Teile genehmigten oder lieferten prima di collegare i piombi di alimentatore a un durch Hoefer, Inc. kann für das Funktionieren, alimentatore.
- Page 6 • Nie działają w buforze temperatury powyżej • Sirkulerer bare vann eller 50/50 vann/ethylene maksymalnego określone specyfikacje techniczne. glykol gjennom oppvarmingen veksleren i så fall Przegrzania spowoduje nieodwracalne szkody dla utstyrer. Ikke forbind oppvarmingen veksleren jednostki! til en vanntapp eller noe kjølemiddelkilde hvor vannet trykket er unregulated. Informações Importantes – • Introduserer Aldri antifreeze eller noe organisk Portuguese løsemiddel inn i noe del av instrumentet. Organiske løsemiddler vil forårsake irreparabel • Se este equipamento é usado numa maneira skade på enheten ! não especificada por Hoefer, Inc. que a • Driver med buffertemperaturer over maksimum protecção fornecida pelo equipamento pode ser ikke spesifiserte teknisk spesifikasjoner. Å comprometida. overoppheting vil forårsake irreparabel skade på • Este instrumento é projectado para uso de interior enheten ! de laboratório só. • Só acessórios e partes aprovaram ou forneceu por Wazne Informacje – Polish Hoefer, Inc. pode ser usada para operar, manter, e servicing este produto. • Jeżeli ten sprzęt jest wykorzystywany w sposób nie określone przez Hoefer, Inc. do ochrony • Só usa um estoque de poder que é CE marcou przewidzianej przez urządzenie może zostać...
- Page 7 Información Importante – • Säkerheten locket måste vara på platsen före koppla kraften tillgången blyen till en kraft tillgång. Spanish • Vänder sig alla kraft tillgång kontroller av och kopplar bort kraften blyen före flytta säkerheten • Si este equipo es utilizado en una manera no locket. especificado por Hoefer, Inc. la protección proporcionado por el equipo puede ser dañada. • Cirkulerar bara vatten eller 50/50 vatten/ethylene glycol genom värmen exchanger i så utrustad fall. • Este instrumento es diseñado para el uso interior Inte kopplar värmen exchanger till en vatten kran del laboratorio sólo. eller något kylmedel källa där vattnet trycket är • Sólo accesorios y partes aprobaron o suministraron unregulated. por Hoefer, Inc. puede ser utilizado para operar, • Inför aldrig kylvätska eller något organiska para mantener, y para atender a este producto. lösningsmedel in i någon del av instrumentet. • Sólo utiliza una alimentación que es CE marcó o Organiskt lösningsmedel ska orsaka irreparable la seguridad certificada por un nacionalmente skada till enheten! reconocido probando el laboratorio. • Använd inte med buffert temperaturer över • La tapa de la seguridad debe estar en el lugar det högsta angivna tekniska specifikationerna.
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Page 8: Waste Electrical And Electronic Equipment (Weee)
Waste Electrical and Electronic Equipment (WEEE) English 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. -
Page 9: Gel Electrophoresis Unit Function And Description
Gel Electrophoresis Unit Function and Description ® The Hoefer SE600 Chroma vertical slab gel electrophoresis unit is 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. -
Page 10: Specifications
CE Certified This declaration of conformity is valid only when the instrument 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 (2) Fig 1. Main components of the Hoefer SE600 Chroma (see Fig 4 for caster components). safety lid Included but not shown: • Gel Seal compound, 1/4 oz. interlock • Spacer-Mate alignment pins template • Glass plates (6) • Wonder Wedge plate separation tool • Buffer dam upper buffer...
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Page 12: Unpacking And Inventory
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. - Page 13 Always install the safety lid before use! Glass plates The SE600 Chroma accommodates 18-cm-wide plates 16 or 8 cm long. Notched divider plates, ordered separately, divide gel sandwiches to form “club sandwiches” of two gels each, so up to four gels can be run at one time.
- Page 14 Clamps Two 16 cm clamps are used to secure the gel sandwich. 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. A laminated gasket in the bottom of each casting cradle seals the bottom of the sandwich when it is clamped into the stand.
- Page 15 Spacers 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 and 2 cm). (May be ordered separately.) Spacer-Mate alignment template This template aligns spacers during sandwich assembly. Combs Combs are available in sizes that form 10, 12, 15, 20, or 28 wells.
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Page 16: Operating Instructions
One to four gels (18 × 16 cm) can be assembled and run in the SE600 Chroma. Both precast gels and self-cast gels can be used. To self-cast multiple gels, kits can be ordered... - Page 17 Construct the gel sandwich and insert into caster Fig 2. Sandwich assembly. Inspect glass plates for nicks. Use only unchipped plates to prevent leaking. Prepare the caster and clamps Place the spirit level into the caster center and adjust spacer clamp glass plates the leveling feet. Loosen all clamp screws and make ridges space for the sandwich by sliding the pressure plates toward the screws.
- Page 18 Club sandwich Fig 3. Club sandwich assembly. A 16-cm-long, notched center-divider plate (ordered Side clamps will accommodate separately) pairs two sandwiches to double the two spacers up to 1.5 mm thick. 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...
- Page 19 Insert a cam into the hole on each side of the casting tray with the ridge (short end) pointing up. Seal the Note: When turning the cams, it is easier to keep the caster gel sandwich against the casting gasket by turning balanced if you turn both toward both cams as far as needed, usually 90°–...
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Page 20: Acrylamide Gels
Acrylamide Gels 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. - Page 21 Overlay each gel with a thin layer of water-saturated 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 one side of the sandwich and allow it to flow across the surface unaided.
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Page 22: Gradient Gels
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... - Page 23 Prepare the monomer solution Calculate the volume of monomer solution needed. Optional: Adjust the higher- percentage acrylamide solution Divide the total volume in half and prepare this to 15% (w/v) sucrose or 25% volume of both the higher- and lower-percentage (v/v) glycerol to improve layering. acrylamide solutions.
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Page 24: Sample Preparation And Loading
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. Sample Preparation and Loading Note: With Coomassie Blue ™... - Page 25 Prepare the sample Increase liquid 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 liquid and dry samples in a test tube. To liquid protein solutions, add an equal volume of 2X buffer.
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Page 26: Final Assembly
Final Assembly Fig 6. Attaching gel sandwiches Upper buffer chamber to the upper buffer chamber. If the assembly leaks, take it to a sink and partially release Rinse both buffer chambers with water and distilled the cams to allow buffer to drain out of the upper chamber. water thoroughly before each use. Disassemble, check alignment Note: Before using the first time, disassemble the of all sandwich components,... - Page 27 stand. Install the cams, ridge pointing down, into the buffer chamber cam holes. Clamp the sandwich in Note: Do not force the cams. place by simultaneously turning one cam clockwise If you encounter unusual and the other counterclockwise a full 180°. resistance, disassemble and inspect clamp and glass alignment along the top of the...
- Page 28 Fig 7. Approximate circulator bath temperature setting. Set the circulator bath temperature setting lower than the desired run temperature by the amount indicated on the graph. This should be checked at three points. power supply setting, W Example: Run parameters: 200 V, 0.05 A (50 mA) 1.
- Page 29 Use the chart (Fig 7, on page 20) 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.
- Page 30 Place the safety lid on the unit by engaging the safety interlock pins before lowering the electrode connections on to the banana plugs. 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.
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Page 31: Separating The Sample
Separating the Sample Electrophoresis parameters for Note: SE600 Chroma unit uses 18-cm-wide plates. The discontinuous polyacrylamide gels gel thickness determines the cross section (and current Gels may be run at either constant current or requirement) for constant constant voltage settings. A constant current current runs. The length of the... - Page 32 The starting voltage for a 1.5 mm slab gel connected to a power supply set to 25 mA is usually 80–90 V (using the SE600 Chroma unit with a Laemmli discontinuous buffer system for SDS gels). The final voltage is typically 250 –...
- Page 33 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.
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Page 34: Care And Maintenance
Care and Maintenance Cleaning Immediately after each use, rinse the upper and lower buffer chambers with water and then • Do not autoclave or heat any rinse thoroughly with distilled water. Handle part above 45 °C. • Do not use organic solvents, the upper buffer chamber with care to prevent abrasives, strong cleaning damaging the banana plug. -
Page 35: 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 36 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 positioning ridges fit inside the grooves. Dirty or damaged Check that the gasket is not damaged or pinched.
- Page 37 problem possible cause remedy Bands are Incomplete gel Degas the stacking-gel solution and avoid trapping air skewed or preparation and bubbles 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 and running gels surface.
- Page 38 problem possible cause remedy Poor band Running Begin electrophoresis as soon as the sample is loaded to resolution conditions 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 reagents.
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Page 39: Bibliography
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). - Page 40 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 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.
- Page 41 O’Farrell, P. H., High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. May 25; 250(10):4007–4021 (1975). Bjellqvist, B., et al., Isoelectric focusing in immobilized pH gradients: principle, methodology and some applications. J. Biochem. Biophys. Methods 6, 317–339 (1982). Görg, A, et al., The current state of two-dimensional electrophoresis with immobilized pH gradients.
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Page 42: Ordering Information
SE6056X for SE600 Chroma High-voltage safety lead set SE6056-HV lower buffer Banana plug, gold, with 2 washers SE6067 chamber SE600 Chroma Heat exchanger/ SE6160 SE6150X lower electrode assembly Glass tube with 2 grommets SE6160-5 for heat exchanger/lower electrode assembly Grommets for heat exchanger/... - Page 43 Gel casters For 1 or 2 gels: Dual Gel Caster, SE6015 basic, 2 gels, 18-cm wide Includes: 2 blank gaskets for 1 or 2 gels. (One included with each SE600 Chroma unit.) For up to 4 gels: Gel Caster Kit, SE675 4 gels, 18 × 16 cm Includes: 8 glass plates, 3 space-saver plates, 5 filler sheets, 100 sheets of wax paper, Spacer-Mate alignment template, and filler plugs.
- Page 44 product qty. code no. Clamps and cams Clamp and Cam Kit, four 16 cm clamps and 8 black cams SE6003UK Replacement thumbscrews for clamps SE6003U-2 Cams, black, for clamps with cam holes SE6005L Clamp assemblies, 16 cm SE6003U Clamp assemblies, 8 cm SE6403U Glass plates 18 × 8 cm Glass plates...
- Page 45 Combs number thickness width of wells (mm) (mm) qty. code no. 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 46 Spacers t hickness (mm) length (cm) width (cm) qty. code no. 0.75 SE6419-2-.75 SE6419-2-1.0 SE6419-2-1.5 0.75 SE6119-2-.75 SE6119-2-1.0 SE6119-2-1.5 SE6118-2-1.0 SE6118-2-1.5 Companion products Hoefer SE100 Plate Mate washing and storage unit SE100 QuickFit connectors, female 3/8" QF3/8 QuickFit connectors, male 3/8" QFX3/8 •...
- Page 47 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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