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

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Authors	HAVLIŠ Jan SHEVCHENKO Andrej MANN Mathias TOMAS Henrik OLSEN Jesper V.
Year of publication	2008
Type	Article in Periodical
Magazine / Source	Nature Protocols
MU Faculty or unit	Faculty of Science
Citation
Field	Genetics and molecular biology
Keywords	ASSISTED-LASER-DESORPTION/IONIZATION; POLYACRYLAMIDE-GELS; NANOELECTROSPRAY; MODEL; SILAC; MS
Description	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.