SDS-PAGE

''The only key step in the procedure is the method of fixing the gel. The gel must be fixed by a non-modifying, precipitation procedure - methanol-acetic acid method is used here. The amounts of solvents used and the times are not generally critical. ''

''Gels should be stained in clean, glass containers. These are separate from those used for Western Blotting.''

''The trays are gently agitated during all steps to ensure even treatment. ''

When possible, remove solutions by aspiration to minimize handling of the gel during the procedure.

Method

 * 1) After electrophoresis, fix the gel by soaking it in the Coomassie Fixing Solution for at least 30 min to ON.
 * 2) Aspirate off the fixing solution and wash the gel at least 3X in ddH2O, at least 10 minutes per wash.
 * 3) Aspirate off the water and cover the gel with Coomassie Stain. You can submerge the gel in excess Coomassie. Stain the gel at room temperature for 3 hours to ON.
 * 4) Aspirate off the Coomassie stain. Destain by soaking the gel in Coomassie Destaining Solution for approximately 1 hour. Longer times may be necessary with gels stained overnight. Placing an absorbent paper on the side helps soak up the dye faster.
 * 5) Transfer the gel onto a protective sheet.
 * 6) Scan the gel and then store gel in container with ddH2O.  The gel can be stored for at least several months at room temperature.