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Mass spectrometric identification of proteins from silver‐stained polyacrylamide gel: A method for the removal of silver ions to enhance sensitivity
Author(s) -
Gharahdaghi Farzin,
Weinberg Catherine R.,
Meagher Denise A.,
Imai Brian S.,
Mische Sheenah M.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990301)20:3<601::aid-elps601>3.0.co;2-6
Subject(s) - mass spectrometry , chromatography , chemistry , matrix assisted laser desorption/ionization , sample preparation in mass spectrometry , sample preparation , protein mass spectrometry , peptide mass fingerprinting , bottom up proteomics , mass spectrum , peptide , desorption , silver stain , ion suppression in liquid chromatography–mass spectrometry , proteomics , tandem mass spectrometry , electrospray ionization , biochemistry , adsorption , microbiology and biotechnology , organic chemistry , biology , gene
Mass 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.