Richert Sophie, Luche Sylvie, Chevallet Mireille, Van Dorsselaer Alain, Leize-Wagner Emmanuelle, Rabilloud Thierry
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR CNRS 7509, ECPM, Strasbourg, France.
Proteomics. 2004 Apr;4(4):909-16. doi: 10.1002/pmic.200300642.
The mechanism by which silver staining of proteins in polyacrylamide gels interferes with mass spectrometry of peptides produced by proteolysis has been investigated. It was demonstrated that this interference increases with time between silver staining and gel processing, although the silver image is constant. This suggested an important role of the formaldehyde used in silver staining development in this interference process. Consequently, a formaldehyde-free staining protocol has been devised, using carbohydrazide as the developing agent. This protocol showed much increased peptide coverage and retained the sensitivity of silver staining. These results were however obtained at the expense of an increased background in the stained gels and of a reduced staining homogeneity.
对聚丙烯酰胺凝胶中蛋白质的银染干扰蛋白质水解产生的肽段质谱分析的机制进行了研究。结果表明，尽管银染图像保持不变，但这种干扰会随着银染与凝胶处理之间的时间增加而增强。这表明在银染显影过程中使用的甲醛在该干扰过程中起重要作用。因此，设计了一种无甲醛染色方案，使用 carbohydrazide 作为显影剂。该方案显示肽段覆盖率大幅提高，并保留了银染的灵敏度。然而，这些结果是以染色凝胶背景增加和染色均匀性降低为代价获得的。
