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PMID: 10217175 Published · ppublish English Journal Article

Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity.

Electrophoresis ·Vol. 20 ·No. 3 ·1999-03-00 ·Pages 601-5

Gharahdaghi F, Weinberg CR, Meagher DA, Imai BS, Mische SM

Abstract

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.

MeSH Terms
Acrylic Resins Electrophoresis, Gel, Two-Dimensional/methods Electrophoresis, Polyacrylamide Gel/methods Ions Proteins/analysis Sensitivity and Specificity Silver Staining/methods Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
Chemicals
Acrylic Resins Ions Proteins polyacrylamide gels
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gharahdaghi F
The Rockefeller University Protein/DNA Technology Center, New York, NY 10021, USA. gharahf@rockvax.rockefeller.edu
Weinberg C R
Meagher D A
Imai B S
Mische S M
Article Info
Journal
Electrophoresis
Abbr.
Electrophoresis
ISSN
0173-0835
Published
1999-03-00
Pages
601-5
Language
English
Region
Germany
NLM ID
8204476
Subset
IM
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