Proteome analysis of polyacrylamide gel-separated proteins visualized by reversible negative staining using imidazole-zinc salts.

作者: Lila Castellanos-Serra , Wilfredo Proenza , Vivian Huerta , Robert L. Moritz , Richard J. Simpson

DOI: 10.1002/(SICI)1522-2683(19990101)20:4/5<732::AID-ELPS732>3.0.CO;2-Q

关键词:

摘要: Identification and characterization of proteins isolated from natural sources by polyacrylamide gel electrophoresis has become a routine technique. However, efficient sample proteolysis subsequent peptide extraction is still problematic. Here, we present an improved protocol for the rapid detection gel-separated proteins, in situ protein modification, proteolytic digestion identification capillary high-performance liquid chromatography/tandem mass spectrometry. This simple technique employs imidazole-zinc reverse stain, in-gel S-pyridylethylation microcrushed pieces with proteases. obviates need buffer exchange or lyophilisation due to all manipulation steps being carried out at near neutral pH thus lends itself readily automation.

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