Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels

A Shevchenko, M Wilm, O Vorm, M Mann - Analytical chemistry, 1996 - ACS Publications
Analytical chemistry, 1996ACS Publications
Proteins from silver-stained gels can be digested enzymatically and the resulting peptides
analyzed and sequenced by mass spectrometry. Standard proteins yield the same peptide
maps when extracted from Coomassie-and silver-stained gels, as judged by electrospray
and MALDI mass spectrometry. The low nanogram range can be reached by the protocols
described here, and the method is robust. A silver-stained one-dimensional gel of a fraction
from yeast proteins was analyzed by nanoelectrospray tandem mass spectrometry. In the …
Proteins from silver-stained gels can be digested enzymatically and the resulting peptides analyzed and sequenced by mass spectrometry. Standard proteins yield the same peptide maps when extracted from Coomassie- and silver-stained gels, as judged by electrospray and MALDI mass spectrometry. The low nanogram range can be reached by the protocols described here, and the method is robust. A silver-stained one-dimensional gel of a fraction from yeast proteins was analyzed by nanoelectrospray tandem mass spectrometry. In the sequencing, more than 1000 amino acids were covered, resulting in no evidence of chemical modifications due to the silver staining procedure. Silver staining allows a substantial shortening of sample preparation time and may, therefore, be preferable over Coomassie staining. This work removes a major obstacle to the low-level sequence analysis of proteins separated on polyacrylamide gels.
ACS Publications