Production and separation of peptides from proteins stained with Coomassie brilliant blue R-250 after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Proteins stained with Coomassie brilliant blue on polyacrylamide gels were digested with lysylendopeptidase in the presence of sodium dodecyl sulfate. Peptide production was similar to that under ordinary conditions of digestion. Peptides were recovered easily and efficiently from the gel pieces and separated by HPLC. The present method for preparation of peptides from proteins separated by sodium dodecyl sulfate gel electrophoresis is quite simple and can be used for sequence analysis of proteins in general at the subnanomolar level.
Kawasaki
H
H
Department of Molecular Biology, Tokyo Metropolitan Institute of Medical Science, Japan.
Emori
Y
Y
Suzuki
K
K
D000595:Amino Acid Sequence; D002135:Calcium-Binding Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D012394:Rosaniline Dyes; D013194:Staining and Labeling
D002135:Calcium-Binding Proteins; D010455:Peptides; D012394:Rosaniline Dyes; C033668:calpastatin; D010450:Endopeptidases; C004692:coomassie Brilliant Blue