Institute of Hygiene and Occupational Medicine, University Hospital Essen, D-45122 Essen, Germany.
Electrophoresis. 2010 Jan;31(4):593-8. doi: 10.1002/elps.200900481.
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 approximately 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.
各种基于 CBB 的方法用于染色 2-D 凝胶电泳分离的蛋白质,比较了它们的灵敏度和分辨率。我们修改了 Kang 的 CBB 染色方案,将磷酸的浓度从原来的 2%改为 8%。事实证明,这是最好的方法。通过采用该方案,可以检测到低至 2ng 的蛋白质量和凝胶中大约 2300 个斑点。修改后的程序执行时间更短。此外,与迄今为止报道的大多数方案相比,该实践更灵敏,能分辨更多的蛋白质斑点,并且与后续的质谱分析兼容。