Journal ArticleDOI
Mass spectrometric identification of proteins from silver‐stained polyacrylamide gel: A method for the removal of silver ions to enhance sensitivity
TLDR
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.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.read more
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Dynamic O-Glycosylation of Nuclear and Cytosolic Proteins CLONING AND CHARACTERIZATION OF A NEUTRAL, CYTOSOLIC β-N-ACETYLGLUCOSAMINIDASE FROM HUMAN BRAIN
TL;DR: It is shown that the transcript is expressed in every human tissue examined but is the highest in the brain, placenta, and pancreas; and cell fractionation suggests that the overexpressed protein is mostly localized in the cytoplasm.
Journal ArticleDOI
A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionization and electrospray ionization- mass spectrometry
Jun X. Yan,Robin Wait,Tom Berkelman,Rachel A. Harry,Jules A. Westbrook,Colin H. Wheeler,Michael J. Dunn +6 more
TL;DR: It is shown that tryptic digests of proteins visualized by the modified stain afford excellent mass spectra by both matrix‐assisted laser desorption/ionization and tandem electrospray ionization, and that the modified silver staining protocol is highly compatible with subsequent mass spectrometric analysis.
Journal ArticleDOI
Silver staining of proteins in polyacrylamide gels
TL;DR: Several variants of silver staining are described here, which can be completed in a time range from 2 h to 1 d after the end of the electrophoretic separation, which is compatible with downstream processing, such as mass spectrometry analysis after protein digestion.
Journal ArticleDOI
Global Profiling of the Cell Surface Proteome of Cancer Cells Uncovers an Abundance of Proteins with Chaperone Function
Bong Kyung Shin,Hong Wang,Anne Marie Yim,François Le Naour,Franck Brichory,Jun Ho Jang,Rong Zhao,Eric Puravs,John Tra,Claire W. Michael,David E. Misek,Samir M. Hanash +11 more
TL;DR: Comprehensive profiling of the cell surface proteome provides an effective approach for the identification of commonly occurring proteins as well as proteins with restricted expression patterns in this compartment.
Journal ArticleDOI
Detection technologies in proteome analysis
TL;DR: New multiplexing capabilities should greatly enhance the applicability of the two-dimensional gel electrophoresis technique with respect to addressing fundamental questions related to proteome-wide changes in protein expression and post-translational modification.
References
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Journal ArticleDOI
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Journal Article
Cleavage of structural proteins during the assemble of the head of bacterio-phage T4
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Journal ArticleDOI
Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels
TL;DR: Silver staining allows a substantial shortening of sample preparation time and may, therefore, be preferable over Coomassie staining, and this work removes a major obstacle to the low-level sequence analysis of proteins separated on polyacrylamide gels.
Journal ArticleDOI
Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry
Matthias Wilm,Andrej Shevchenko,Tony Houthaeve,Stephen Breit,Lothar Schweigerer,Lothar Schweigerer,Theodore Fotsis,Matthias Mann +7 more
TL;DR: A simple and robust technique for the sequencing of proteins isolated by polyacrylamide gel electro-phoresis, using nano-electrospray3,4 tandem mass spectrometry5,6 and multiple-sequence stretches of up to 16 amino acids are obtained.
Journal ArticleDOI
Rapid identification of proteins by peptide-mass fingerprinting
TL;DR: Peptide-mass fingerprints can prove as discriminating as linear peptide sequences, but can be obtained in a fraction of the time using less protein, which allows for a rapid identification of a sample protein before committing it to protein sequence analysis.