Electrophoresis Grade
Most commonly used stain in protein electrophoresis. A fast acting, sensitive dye which can be used on SDS gels, IEF gels, and standard gels.
Typical Coomassie Solution:
Prepare in distilled, deionized water:
50% (v/v) methanol
0.05% (w/v) Coomassie Brilliant Blue R-250
10% acetic acid
40% Water
Dissolve the Coomassie Brilliant Blue R in methanol before adding acetic acid and water (filtration is not generally needed). Solution can be stored for 6 months. If precipitate is observed following prolonged storage, filter to obtain a homogeneous solution.
General Protein Staining Procedure:1
Cover the polyacrylamide gel with 3 to 5 volumes of a fixing solution (50% [v/v] methanol, 10% [v/v] acetic acid; 40% water). Agitate slowly 2 hours on an orbital shaker.
Remove the fixing solution and cover the gel with the Coomassie Brilliant Blue solution from above. Slowly agitate on the orbital shaker for 4 hours.
Remove the staining solution and rinse the gel briefly with fixing solution.
Remove the fixing solution and cover the gel with a destaining solution (7% acetic acid; 5% methanol; 88% water). Slowly agitate on the orbital shaker for 2 hours. Remove the destaining solution. Repeat with fresh destaining solution until blue bands and a clear background appear.
The gel can then be stored in 7% acetic acid, water, or dried for storage.
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