Assessing factors for reliable quantitative proteomics based on two-dimensional gel electrophoresis

Proteomics
Julie FiévetDominique de Vienne

Abstract

We statistically analysed various factors to get accurate estimates of protein quantities from two-dimensional gels. Yeast proteins were labelled with (35)S or stained with Coomassie Brilliant Blue G-250, and spots were automatically quantified with software packages Kepler, ImageQuaNT, Melanie 3.0 and Progenesis. The different software packages proved to have very similar performances. With (35)S-labelled actin spot as a reference, we studied the staining efficiency of colloidal Coomassie blue as a function of amino acid composition of the protein, and derived an equation to estimate the number of molecules per cell from blue-stained proteins. Absolute quantification of most glycolytic enzymes was carried out in two yeast strains.

Citations

Dec 29, 2012·Molecular & Cellular Proteomics : MCP·Warren AlbertinDelphine Sicard
Apr 13, 2010·Molecular & Cellular Proteomics : MCP·Natasha A KarpKathryn S Lilley
Sep 28, 2006·Clinical and Experimental Pharmacology & Physiology·Kym J GuelfiPeter G Arthur
Nov 17, 2009·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Julie B FiévetDominique de Vienne
May 18, 2013·Archives of Physiology and Biochemistry·Anton PoschStefan Lehr
Mar 27, 2015·Environmental Microbiology·Victor SabarlyChristine Dillmann
Sep 15, 2009·Proteomics·Peggy Baudouin-CornuJean Labarre
Jun 15, 2007·Free Radical Biology & Medicine·Mikael MolinJean Labarre
Jul 30, 2009·Proteomics·Jérôme DenardFedor Svinartchouk
Jul 1, 2010·Journal of Biomedicine & Biotechnology·Raquel González-FernándezJesús V Jorrín-Novo
Nov 16, 2004·Proteomics·Angelika GörgMichael J Dunn
Sep 26, 2007·Proteomics·Natasha A Karp, Kathryn S Lilley
Jan 16, 2007·Protein Engineering, Design & Selection : PEDS·Russell E LyonsChristopher M Elvin
Oct 24, 2007·Journal of Proteome Research·Cristina-Maria Valcu, Mihai Valcu
Oct 11, 2005·Journal of Proteome Research·Natasha A KarpKathryn S Lilley

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