Abstract

Proteins from silver-stained gels can be digested enzymatically and the resulting peptide analyzed and sequenced by mass spectrometry. Standard proteins yield the same peptide maps when extracted from Coomassie- and silver-stained gels, as judged by electrospray and MALDI mass spectrometry. The low nanogram range can be reached by the protocols described here, and the method is robust. A silver-stained one-dimensional gel of a fraction from yeast proteins was analyzed by nano-electrospray tandem mass spectrometry. In the sequencing, more than 1000 amino acids were covered, resulting in no evidence of chemical modifications due to the silver staining procedure. Silver staining allows a substantial shortening of sample preparation time and may, therefore, be preferable over Coomassie staining. This work removes a major obstacle to the low-level sequence analysis of proteins separated on polyacrylamide gels.

Keywords

ChemistryChromatographySilver stainMass spectrometryCoomassie Brilliant BlueStainingProtein mass spectrometryTandem mass spectrometryPolyacrylamidePolyacrylamide gel electrophoresisPeptideProteomicsBiochemistryMolecular biologyEnzyme

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Year
1996
Type
article
Volume
68
Issue
5
Pages
850-858
Citations
9002
Access
Closed

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Andrej Shevchenko, Matthias Wilm, Ole Vorm et al. (1996). Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels. Analytical Chemistry , 68 (5) , 850-858. https://doi.org/10.1021/ac950914h

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DOI
10.1021/ac950914h