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Summary of Contents for Hoefer SE300 miniVE
- Page 1 Hoefer SE300 miniVE Electrophoresis and Electrotransfer Unit SE300-IM...
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Page 2: Table Of Contents
Contents Important Information ........... ii Waste Electrical and Electronic Equipment (WEEE) .......vii Introduction ............1 Unpacking ............2 Specifications ............3 The Electrophoresis module ........4 Electrophoresis ............14 Care and maintenance .........15 Electrophoresis troubleshooting ......16 The blot module ..........18 Blotter care and maintenance .......23 Blotter troubleshooting .........24 Bibliography ............26 Ordering information ..........27... -
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. - 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...
- 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 labora- des dommages irréparables à l’unité ! torio interno solo. • Solo gli accessori e le parti hanno approvato o Wichtige Informationen – German 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 o la gestellte Schutz verschlechtert werden. sicurezza certificato da un nazionalmente ricon- • Dieses Instrument wird für den Innenlaborge- osciuto testando il laboratorio. brauch nur dafür entworfen. • Il coperchio di sicurezza deve essere nel luogo • Nur Zusätze und Teile genehmigten oder lieferten...
- 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-...
- 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 –...
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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: Introduction
Introduction ® The Hoefer SE300 miniVE vertical electropho- Note: Minimum power supply ratings: 50 mA, 250 V constant resis system performs vertical gel electrophoresis current or constant voltage. on mini-format gels. The basic unit includes two electrophoresis modules. Each module holds one gel sandwich, 10 cm wide and up to 10.5 cm... -
Page 10: Unpacking
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 tran- sit. -
Page 11: Specifications
Hoefer, Inc. except for alterations described in the user manual, and connected to other CE labeled instru- ments or products recommended or approved by Hoefer, Inc. -
Page 12: The Electrophoresis Module
The Electrophoresis module This section describes the use of the electropho- resis module. For instructions on using the blot module, see page 18. The electrophoresis module accepts both self- cast and precast gels 8 cm wide, from 8–10.5 cm long, and 0.75–1.5 mm thick. For instructions on using the module with precast gels, see page 10. - Page 13 Lay the module flat on a work surface. Preparing self-cast gels One single gel can be cast on the module. To cast several gels, use a 4-gel caster such as the Hoefer SE235 caster (see Ordering information on page 27). Assemble the gel sandwich ...
- Page 14 Place and seal the sandwich on the module Important! Proper alignment is Take care to “square” the three sealing sides of the essential to prevent leakage. sandwich. Hold the sandwich like a deck of cards and gently tap the bottom against a flat surface. Note: Once the sandwich is care- ...
- Page 15 If the spacers and glass plates are not perfectly aligned against the stops, use the stiff end of the Hoefer Wonder Wedge to press against the edges of the spacer and glass plates and position them flush against the guide foot.
- Page 16 Lock the sealing plate into the closed position by engaging each tab in its topmost notch (Fig 9). Fig 9. Assembled module, with tabs engaged in top notch tabs engaged in top notch. Note: To test for alignment, pass ...
- Page 17 A 1-cm stacking gel below the wells Fill to 3 cm below the top of the rectangular glass plate. Overlay each gel with a thin layer of water-saturated n-butanol, water, or diluted gel buffer to prevent exposing the monomer solu- tion to oxygen.
- Page 18 Insert a comb (at a slight angle to prevent trapping Note: If the gel has wells, skip to air) into the sandwich, allowing the comb sides to rest “Final assembly” on page 11. on the spacers. Tip: To calculate the volume, measure the distance in centime- Allow a minimum of one hour for the gel to polymerize.
- Page 19 Fig 11. Securing the cassette. Check that both gel surfaces will contact buffer. Check that the bottom gel-contact slot is exposed. Move the sealing plate into the “half open” position to prepare for electrophoresis. Apply gentle inward pres- sure to both tabs and lock them into the lower notch.
- Page 20 Fig 12. Preparing for electrophoresis. Tip: As an aid in loading samples, mark the well location with a labo- ratory marking pen or use locating decal. The locating decal only works with miniVE combs and not precast gels. the half-open postion for electrophoresis upper buffer chamber The minimum and maximum levels are marked.
- Page 21 Underlay the sample into the wells using a micro- pipet or fine-tipped microsyringe. Table 1 shows the Note: The amount of protein volume of sample required for different numbers of sample added to each well wells and comb thicknesses. depends on both the sensitivity of the staining method and the distri- bution of protein among separate Table 1:...
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Page 22: Electrophoresis
Remove the gel from the sandwich or cassette. Gently loosen and then slide away both spacers. Slip an extra spacer or the Hoefer Wonder Wedge into the bottom edge to prevent breaking the “ears” of the notched plates and separate the plates. -
Page 23: Care And Maintenance
Carefully lift the gel from the plate and lay it into a tray containing stain, fixative, or transfer buffer. Clean the unit as described in “Care and mainte- nance” below. Care and maintenance • Do not autoclave or heat any part above 75 °C. •... -
Page 24: Electrophoresis Troubleshooting
Electrophoresis Troubleshooting problem solution Smile effect on the buffer front To reduce the running temperature: • Fill the tank to the maximum (marked) buffer level. • Prechill the buffer. • Decrease the current or voltage setting. (10 mA per 0.75 mm gel, 15 mA per 1.5 mm thick gel.) •... - Page 25 problem solution Stained sample collects: 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.
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Page 26: The Blot Module
The blot module The Hoefer miniVE Blot Module, which can be ordered separately, performs electrotransfers on mini-format gels. Each module holds up to two gels, 8.2 cm wide and up to 10.4 cm long. One or two modules can be run at the same time. - Page 27 Preparation Optional: Passive cooling Chill approximately 2 liters of deionized water to 4 °C. (Filling the tank with chilled water serves as a heat sink during electrotransfer.) Prepare transfer buffer Stack assembly requires approximately 250 ml transfer buffer and an additional 300 – 350 ml buffer is required to fill each module.
- Page 28 Prepare the transfer stack Transfer the sample as soon as possible after electrophoresis to minimize sample diffusion within the gel. Electrophoretic transfer can be performed on as many as four mini gels at one time, if two gels are placed in each of two modules.
- Page 29 Assemble the transfer stack so that molecules will migrate to the membrane (Fig 15). For negatively charged macromolecules—such as Fig 15. Assembling the transfer proteins run in an SDS gel and nucleic acids— stack. assemble the transfer stack on the black (cathode) side.
- Page 30 Check the position of the transfer stack. The transfer stack should be centered on the elec- trode plate. No layer should be pinched when the module is closed. Fold the empty half of the cup over the stack and press the halves together to snap the module closed.
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Page 31: Blotter Care And Maintenance
Electrotransfer Electrophoretic transfer conditions for blotting Important! Buffer conductivity proteins in Towbin buffer: 25 V for 1–2 hours, increases with increasing temperature, providing a 300 – 400 mA. positive feedback that results in rapid heating. We recommend After electrotransfer programming the power supply to hold the current setting constant ... -
Page 32: Blotter Troubleshooting
Blotter troubleshooting problem possible cause solution Incomplete Blank areas on Remove all trapped air bubbles in the transfer stack; take espe- transfer the membrane cially great care during stack assembly to prevent air bubbles from forming as each layer is placed. Check electrode continuity. - Page 33 problem possible cause solution Inefficient Chemical Fix or crosslink the mole to the requirements of the nucleic acid, binding parameters protein, or membrane type. Prepare protein transfer buffer without SDS. SDS can improve transfer efficiency but reduces binding. Verify the optimal amount of methanol required for the membrane type and check the buffer solution.
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Page 34: Bibliography
Bibliography Polyacrylamide gel electrophoresis Adams, L.D. and Gallagher, S.R. 1992. Two-Dimensional Gel Electrophoresis Using the O’Farrell System. Current Protocols in Molecular Biology, 10.4.1–10.4.13. Gallagher, S.R., and Smith, J.A. 1991. Electrophoretic separation of proteins. Current Protocols in Molecular Biology. (F.A. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. -
Page 35: Ordering Information
Ordering information product quantity code no. Hoefer SE300 miniVE, complete SE300-10A-1.0 Includes Basic unit, 3 rectangular glass plates, 3 notched plates, 2 each 1.0 mm thick, 10 well combs and 1.0 mm thick spacer sets. Blot Module SE302 Includes 3 Dacron ™... - Page 36 SE3005 Blotting paper, 9 –10.5 cm 50/pk TE26 Gel casters Hoefer SE235 4-Gel Caster, 2 to 4 gels, 10 × 10.5 cm SE235 Power supplies PS200HC Power Supply, 200 V, 2000 mA, 200 W PS200HC PS300B Power Supply, 300 V, 500 mA, 90 W...
- Page 37 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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