Application of Heat to Quickly Stain and Destain Proteins Stained with Coomassie Blue

Abstract

Proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis have been visualized reliably by staining with Coomassie Brilliant Blue. In this chapter, we show that it is possible to drastically reduce protein staining and destaining time, while simultaneously increasing detection sensitivity, with the application of heat. It took 5 min to stain proteins at 55, 62.5, or 70 °C for a 1.5 mm gel, while it took 45, 45, and 20 min respectively for destaining. The time for staining was 1 min for a 0.8 mm gel at 65 °C, 2 min at 60 °C and 5 min at 55 °C. The destaining of proteins separated on a 0.8 mm gel took 8, 15, and 20 min at 65, 60, and 55 °C respectively. Proteins can be stained and destained rapidly with the use of heat, while enhancing detection sensitivity.

Keywords: Coomassie Brilliant Blue; Destaining; Heat; SDS-PAGE.