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Silver‐ and Coomassie‐staining protocols: Detection limits and compatibility with ESI MS
Author(s) -
Winkler Christiane,
Denker Katrin,
Wortelkamp Stefanie,
Sickmann Albert
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600670
Subject(s) - staining , silver stain , coomassie brilliant blue , chemistry , chromatography , bovine serum albumin , microbiology and biotechnology , mass spectrometry , biology , genetics
Staining protocols for PAGE have to be sensitive and should not impair further MS analysis of selected samples. In this study, the MS compatibility of different silver‐ and Coomassie‐staining protocols with a nano‐LC‐MS/MS system was systematically elucidated. Altogether, 13 different silver‐staining, 1 imidazole‐staining and 2 Coomassie‐staining protocols were used and compared to each other for their achieved sequence coverage and their detection sensitivity. Three proteins were used as model proteins (bovine serum albumin, rabbit L ‐lactate dehydrogenase, bovine milk β‐lactoglobulin) in decreasing concentration (12 pmol down to 30 fmol) and different staining protocols were applied. The conclusion of this study is that two silver‐staining protocols (Blum, H. et al.,. Electrophoresis 1987, 8 , 93–99 and Shevchenko, A. et al.,. Anal. Chem. 1996, 68 , 850–858) combine good sequence coverage and good sensitivity and are recommended for nano‐LC‐MS/MS analysis.