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Summary of Contents for Hoefer SE 260
- Page 1 SE 260 Hoefer SE 260 mini-vertical gel electrophoresis unit SE260-IM/Rev. A0/05-04...
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Page 3: Table Of Contents
Page finder 1. Gel Electrophoresis Unit Function ... 1 and Description 2. Important information ....4 3. - Page 4 •...
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Page 5: Gel Electrophoresis Unit Function
45 minutes, and only a minimal amount of sample is required. The SE 260 accommodates one or two 10 × 8 cm or 10 × 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. - Page 6 • used in laboratory locations, Max. amperage 500 mA • used as delivered from Max. temperature 45 °C Hoefer, Inc. except for alter- Environmental Indoor use: 4–40 °C ations described in the user operating conditions Humidity up to 80% manual, and Altitude up to 2000 m •...
- Page 7 Color-coded leads (2) SE6056-HV Safety lid SE256 •...
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Page 8: Important Information
• If running only one gel, block off the unused part of the core with a glass plate. Do not fill this side with buffer. • Only accessories and parts approved or supplied by Hoefer, Inc. may be used for operating, maintaining, and servicing this product. •... - Page 9 Ne pas remplir le côté vide avec du tampon. • Seulement les accessoires et piéces detachées approuvés ou fournis par Hoefer, Inc. sont recom- mandés pour l’utilisation, l’entretien et réparation de cet appareil.
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Page 10: Operating Instructions
3.1 Prepare the gel sandwich Both precast and self-cast gels can be run in the SE 260 units. For longer gel length, 10 × 10.5 cm plates, gels can be cast in the Hoefer SE 235 4-gel caster. The SE 260 can also accommodate shorter length gel in 10 ×... - Page 11 3.2 Prepare the unit 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.
- Page 12 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.
- Page 13 3.3 Place the gel sandwich Fig. 3a–b. Gel sandwich installation. Rinse away the overlay with distilled water and drain any excess water. If installing a self-cast or precast 10 × 8 cm gel sand- wich, align the bottom of the plate with the bottom of the core.
- Page 14 Clamp the sandwich in place 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.
- Page 15 3.4 Sample preparation and loading 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: Note: Stacking gel resolution measure the distance, in cm, from the top of the is optimal when poured just resolving gel to the notch in the alumina plate.
- Page 16 Fill the sample wells and each upper buffer cham- ber that will be used with running buffer. One upper buffer chamber holds approximately 75 mL. Underlay the sample into the wells using a fine-tipped Note: The amount of protein microsyringe.
- Page 17 3.5 Final assembly Fill the lower buffer chamber with running buffer. The SE 260 holds about 250 mL. Check that the lower electrode (running along the bottom of the the upper buffer chamber core) is completely submerged. Note: If using precast gels, check that the lower gel/ buffer contact surface is exposed (the colored plastic tape must be removed.)
- Page 18 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 of electrophoretic migra- tion remains unchanged throughout the run. Under these conditions, voltage increases as the run proceeds.
- Page 19 Gently 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.
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Page 20: Care And Maintenance
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. -
Page 21: Troubleshooting
5. Troubleshooting problem solution Smile effect on the buffer front 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. - Page 22 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 23: Appendix
Commonly, the amount of crosslinker used (% C) is 2.6%. In the follow- ing example system, the resolving gel compo- SE 260 results: sition is 10% T, 2.6% C, which results in a Lane 1: SDS-6H, high MW medium pore size. - Page 24 Final concentrations separating gel stacking gel electrophoresis buffer Acrylamide conc. 10% T*, 2.6% C 4% T, 2.6% C Tris-Cl 0.375 M 0.125 M Tris-glycine 0.025 M Tris base 0.192 M glycine ~8.3 0.1% 0.1% 0.1% Ammonium persulfate (APS) 0.05% w/v 0.05–0.1% w/v †...
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Page 25: Bibliography
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 sepa- ration of proteins. Current Protocols in Molecular Biology. (F.A. Ausubel, R. Brent, R.E. Kingston, D.D. - Page 26 •...
- Page 27 Wonder Wedge plate separation tool SE1514 High voltage safety lead set SE6056-HV Gel Seal (0.25 oz) SE6070 Spring clamps, for SE 260 and gel casters SE252 Well locating label SE212 Glass and Alumina Plates 10 × 8 cm Notched alumina plates...
- Page 28 product quantity code number Spacers thickness (mm) length (cm) 0.75 SE2119T-2-.75 1.00 SE2119T-2-1.0 1.50 SE2119T-2-1.5 0.75 10.5 SE2619T-2-.75 1.00 10.5 SE2619T-2-1.0 1.50 10.5 SE2619T-2-1.5 Teflon Combs well thickness (mm) width (mm) 0.75 13.0 SE211A-5-.75 1.00 13.0 SE211A-5-1.0 1.50 13.0 SE211A-5-1.5 1.00 SE211A-9-1.0 0.75...
- Page 29 100 sheets wax paper, space saver plate, 5 filler sheets, set of filler plugs and Spacer-Mate assembly template.(Order combs and spacers separately.) Power Supplies Hoefer EPS 2A200 PSA2A200 Miscellaneous Hoefer SE 100 PlateMate washing and storage unit SE100 TE 22 Transphor Tank Unit TE22 QuickFit connector, male, 3/8” QFX3/8...
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- Page 32 Printed in the USA Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA www.hoeferinc.com...
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