In-gel digestion for mass spectrometric characterization of proteins and proteomes.
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.
Shevchenko
Andrej
A
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany. shevchenko@mpi-cbg.de
Tomas
Henrik
H
Havlis
Jan
J
Olsen
Jesper V
JV
Mann
Matthias
M
D004586:Electrophoresis; D013058:Mass Spectrometry; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
D011506:Proteins