title: Silver- and Coomassie-staining protocols: detection limits and compatibility with ESI MS.
abstract: 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 beta-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.
authors.affiliation: Rudolf-Virchow-Center, DFG-Research Center for Experimental Biomedicine, University of Würzburg, Würzburg, Germany.
authors.forename: Christiane
authors.initials: C
authors.lastname: Winkler
authors.forename: Katrin
authors.initials: K
authors.lastname: Denker
authors.forename: Stefanie
authors.initials: S
authors.lastname: Wortelkamp
authors.forename: Albert
authors.initials: A
authors.lastname: Sickmann
mesh_terms: D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D003720:Densitometry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D010449:Peptide Mapping; D011506:Proteins; D040901:Proteomics / Q000379:methods; D011817:Rabbits; D012394:Rosaniline Dyes / Q000737:chemistry*; D012834:Silver / Q000737:chemistry*; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D011506:Proteins; D012394:Rosaniline Dyes; D012834:Silver; C004692:coomassie Brilliant Blue