CBB staining protocol with higher sensitivity and mass spectrometric compatibility

Various CBB-based methods for staining proteins separated by 2-D gel electrophoresis were compared with regard to sensitivity and resolution. A modified Kang's CBB staining protocol, which we have modified, includes phosphoric acid in a concentration of 8% instead of the original 2%. This proved to be the best approach. Protein amounts as low as 2 ng and similar to 2300 spots in the gel can be detected by employing this protocol. The modified procedure takes less time to carry out. Moreover, this practice is more sensitive and resolves more protein spots than most protocols reported to date and is compatible with subsequent mass spectrometric analysis.