In-gel Digestion for Mass Spectrometric Characterization of Proteins and Proteomes

Nat Protoc. 2006;1(6):2856-60. doi: 10.1038/nprot.2006.468.

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.

MeSH terms

  • Electrophoresis
  • Mass Spectrometry
  • Proteins / metabolism*
  • Proteomics / methods*

Substances

  • Proteins