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Methods Mol Biol. 2005;310:227-40.

Peptide mass fingerprinting: protein identification using MALDI-TOF mass spectrometry.

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  • 1Proteomics Research Group, Babraham Institute, Cambridge, UK.

Abstract

Matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF)-mass spectrometry (MS) is now routinely used in many laboratories for the rapid and sensitive identification of proteins by peptide mass fingerprinting (PMF). We describe a simple protocol that can be performed in a standard biochemistry laboratory, whereby proteins separated by one- or two-dimensional gel electrophoresis can be identified at femtomole levels. The procedure involves excision of the spot or band from the gel, washing and de-staining, reduction and alkylation, in-gel trypsin digestion, MALDI-TOF MS of the tryptic peptides, and database searching of the PMF data. Up to 96 protein samples can easily be manually processed at one time by this method.

PMID:
16350956
[PubMed - indexed for MEDLINE]
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