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


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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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).

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