Two-Dimensional Peptide Mapping of Protease-Digested Proteins from Two-Dimensional Electrophoresis Gel Spots

摘要

A simple procedure is described to extract proteins from polyacrylamide gels and subsequently separate them after partial proteolysis. This method is useful for the biochemical characterization of individual proteins in two-dimensional gel patterns and for the identification of both similarities and differences between a pair of related protein spots. Because the number of gel spots to be extracted and the extraction volume can be adjusted in this procedure, differences in the primary structure of both abundant and minor proteins can be investigated. By extracting protein from 10 or more gel spots, enough protein should be obtained to allow amino acid sequencing of peptides that carry significant amino acid differences. 7 refs., 1 fig., 1 tab. (ERA citation 12:016795)