Document Detail


Qualitative and quantitative proteomics by two-dimensional gel electrophoresis, peptide mass fingerprint and a chemically-coded affinity tag (CCAT).
MedLine Citation:
PMID:  14651868     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The chemically-coded affinity tag (CCAT) method combines standard electrophoresis protocols with MALDI-TOF-MS analysis to identify and quantify protein abundances in complex samples in one step. This method is designed to fit into the workflow of SDS-PAGE or two-dimensional electrophoresis (2-DE) only requiring basic proteome laboratory equipment. Prior to electrophoresis two protein samples are separately labelled with a heavy or a light version of the CCAT reagent via reduced cysteines in the proteins. Equal amounts are then combined and electrophoretically separated. Proteins can then be excised from the gel to obtain their peptide mass fingerprint by mass spectrometry. This fingerprint enabled not only identification, but also quantification by comparing relative peak intensities of CCAT-labelled peptides. In this article, we display how the CCAT method can be used to analyse two protein samples in one gel and that the peak intensities of labelled peptides reflect the abundance of a protein in it.
Authors:
Steven Alexander Watt; Thomas Patschkowski; Jörn Kalinowski; Karsten Niehaus
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't; Validation Studies    
Journal Detail:
Title:  Journal of biotechnology     Volume:  106     ISSN:  0168-1656     ISO Abbreviation:  J. Biotechnol.     Publication Date:  2003 Dec 
Date Detail:
Created Date:  2003-12-03     Completed Date:  2004-08-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8411927     Medline TA:  J Biotechnol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  287-300     Citation Subset:  IM    
Affiliation:
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, POB 100131, 33501 Bielefeld, Germany.
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MeSH Terms
Descriptor/Qualifier:
Affinity Labels / chemistry
Bacterial Proteins / analysis*,  chemistry*
Electrophoresis, Gel, Two-Dimensional / methods*
Peptide Mapping / methods*
Proteomics / methods*
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
Xanthomonas campestris / metabolism
Chemical
Reg. No./Substance:
0/Affinity Labels; 0/Bacterial Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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