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Cited by 8,995

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Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels

Shevchenko, Andrej, Wilm, Matthias, Vorm, Ole, Mann, Matthias·1996·Analytical Chemistry

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.

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Cited by

Metadata

Authors
Shevchenko, A., Wilm, M., Vorm, O., & Mann, M.
Venue
Analytical Chemistry
Published
1996-03-01
Volume
68
Issue
5
Pages
850-858
ISSN
  • 0003-2700 (ISSN-L)
  • 1520-6882
OpenAlex
W2078594804
Cited by
8,995
References
2
FWCI
58.14
Type
journal article
Language
en
Open access
closed

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