Heat mediated quick Coomassie blue protein staining and destaining of SDS-PAGE gels.
For the detection of proteins on sodium dodecyl sulphate-polyacrylamide gel electrophoresis, Coomassie blue is used commonly on account of its simplicity and reliability. In this report we show that enhanced heat, in addition to dramatically decreasing the time required for staining and destaining, also significantly increased the detection sensitivity. For a 1.5 mm gel, the staining time was 5 min at 55, 62.5 or 70 degrees C while the destaining time was 45, 45 and 20 min respectively. For a 0.8 mm gel, the staining time could be reduced to 1 min at 65 degrees C compared to 2 min at 60 degrees C and 5 min at 55 degrees C. The destaining time required was 8, 15 and 20 min at the respective temperatures.
Arthritis and Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City 73104, USA. kurienb@omrf.ouhsc.edu
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D004396:Coloring Agents / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006358:Hot Temperature; D012394:Rosaniline Dyes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors
D004396:Coloring Agents; D012394:Rosaniline Dyes; C048139:Coomassie blue