title: Peptide mass fingerprinting: protein identification using MALDI-TOF mass spectrometry.
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.
authors.affiliation: Proteomics Research Group, Babraham Institute, Cambridge, UK.
authors.forename: Judith
authors.initials: J
authors.lastname: Webster
authors.forename: David
authors.initials: D
authors.lastname: Oxley
mesh_terms: D030562:Databases, Protein; D006801:Humans; D008970:Molecular Weight; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D010455:Peptides; D011506:Proteins