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

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

Nature protocols (2007-04-05)
Andrej Shevchenko, Henrik Tomas, Jan Havlis, Jesper V Olsen, Matthias Mann
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.

MATERIALS
Product Number
Brand
Product Description

Millipore
Seppro Ammonium Bicarbonate Buffer
Sigma-Aldrich
Ammonium bicarbonate, puriss., meets analytical specification of Ph.Eur., BP, E 503, 99-101%