Nature Protocols 1, - 2856 - 2860 (2007) doi:10.1038/nprot.2006.468

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

Andrej Shevchenko¹, Henrik Tomas¹, Jan Havlis breve¹, Jesper V Olsen², Matthias Mann²

1. Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
2. Max Planck Institute for Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.

 

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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