In-gel digestion for mass spectrometric characterization of proteins and proteomes
Nature protocols, 2006•nature.com
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.
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.
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.
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