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In-gel digestion for mass spectrometric characterization of proteins and proteomes

Nature Protocols volume 1, pages 2856–2860 (2006)Cite this article

Abstract

In-gel digestion of proteins isolated by gel electrophoresis is a cornerstone of mass spectrometry (MS)-driven proteomics. The 10-year-old recipe by Shevchenko et al. has been optimized to increase the speed and sensitivity of analysis. The protocol is for the in-gel digestion of both silver and Coomassie-stained protein spots or bands and can be followed by MALDI-MS or LC-MS/MS analysis to identify proteins at sensitivities better than a few femtomoles of protein starting material.

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Figure 1: Characterization of a modified protein by GeLCMS.
Figure 2: Characterization of a complex proteome.

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Authors and Affiliations

  1. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, Dresden, 01307, Germany

    Andrej Shevchenko, Henrik Tomas & Jan Havli

  2. Max Planck Institute for Biochemistry, Am Klopferspitz 18, Martinsried, 82152, Germany

    Jesper V Olsen & Matthias Mann

Authors
  1. Andrej Shevchenko
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  2. Henrik Tomas
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  3. Jan Havli
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  4. Jesper V Olsen
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Correspondence to Andrej Shevchenko or Matthias Mann.

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Shevchenko, A., Tomas, H., Havli, J. et al. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1, 2856–2860 (2006). https://doi.org/10.1038/nprot.2006.468

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