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

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 Co...

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Veröffentlicht in:Nature protocols 2007-01, Vol.1 (6), p.2856-2860
Hauptverfasser: Shevchenko, Andrej, Mann, Matthias, Tomas, Henrik, Havli, Jan, Olsen, Jesper V
Format: Artikel
Sprache:eng
Schlagworte:
Acids
Analytical Chemistry
Biological Techniques
Biomedical and Life Sciences
Computational Biology/Bioinformatics
Electrophoresis
Enzymes
Life Sciences
Mass Spectrometry
Methods
Microarrays
Molecular weight
Organic Chemistry
Peptides
Proteins
Proteins - metabolism
Proteolysis
Proteomics
Proteomics - methods
Protocol
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Zusammenfassung: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.
ISSN:1754-2189
1750-2799
DOI:10.1038/nprot.2006.468