Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: A method for the removal of silver ions to enhance sensitivity
1999; Wiley; Volume: 20; Issue: 3 Linguagem: Inglês
10.1002/(sici)1522-2683(19990301)20
ISSN1522-2683
AutoresFarzin Gharahdaghi, Catherine Weinberg, Denise A. Meagher, Brian S. Imai, Sheenah M. Mische,
Tópico(s)Ion-surface interactions and analysis
ResumoMass spectrometry is a powerful technique for the identification of proteins at nanogram quantities. However, some degree of sample preparation prior to mass spectrometry is required, and silver-stained protein gel samples are most problematic. Here we report our strategy to obtain peptide mass profiles from silver-stained protein gel samples from one- or two-dimensional gels by destaining prior to enzymatic digestion. This study demonstrates that by using the destaining method, the sensitivity and quality of mass spectra is increased for matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometric analysis, permitting more proteins to be identified by peptide mass database analysis.
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