Contents Important Information ..........ii Waste Electrical and Electronic Equipment (WEEE) .......vii 1. Gel Electrophoresis Unit Function and Description ........1 2. Specifications ...........2 3. Operating Instructions ........4 4. Care and Maintenance ........14 5. Troubleshooting ..........15 Appendix ............17 Bibliography ............19 Ordering Information ..........20 •...
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 • The safety lid must be in place before connecting jednotka! the power supply leads to a power supply. Vigtig Information – Danish • Turn all power supply controls off and disconnect...
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.
45 minutes, and only a minimal amount of sample is required. The SE250 accommodates one or two 10 × 8 cm gel sandwiches. The upper buffer chamber is formed when the notched side of a gel sandwich is sealed against the silicone rubber gasket.
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.
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Fig 1. Main components Color-coded leads (2) SE250 electrophoresis unit. SE6056-HV Included but not shown: Safety lid Glass plates SE256 Notched alumina plates Gel seal, 1/4 oz. Well-locating decal Required but not included: Upper buffer chamber core Power supply with a...
3.1 Prepare the gel sandwich Both precast and self-cast gels can be run in the Note: All electrophoresis SE250 units. This unit accepts gels in 10 × 8 cm accessories and kits are listed in plates, which can be cast in a Hoefer SE215, the the ordering section.
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.
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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.
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3.3 Place the gel sandwich Rinse away the overlay with distilled water and drain any excess water. If installing a self-cast or precast 10 × 8 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 3).
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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.
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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 is optimal when poured just before measure the distance, in cm, from the top of the electrophoresis.
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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.
Note: If using precast gels, Fill the lower buffer chamber with running buffer. The check that the lower gel/buffer SE250 holds about 150 mL. Check that the lower contact surface is exposed. electrode (running along the bottom of the the upper buffer chamber core) is completely submerged.
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 migration remains unchanged throughout the run. Under these conditions, voltage increases as the run proceeds.
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.
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.
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.) •...
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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.
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 SE250 results: composition is 4% T, 2.6% C. The % T in the Lane 1: SDS-6H, high MW stacking gel is lower because a larger pore size is ™...
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Final concentrations separating gel stacking gel electrophoresis buffer Acrylamide concentration 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 TEMED †...
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.
Wonder Wedge plate separation tool SE1514 High voltage safety lead set SE6056-HV Gel Seal (0.25 oz.) SE6070 Spring clamps, for SE250 and gel casters SE252 Well locating label SE212 Glass and Alumina Plates 10 × 8 cm Notched alumina plates...
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