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Proteomics. 2004 Jul;4(7):1939-49.

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

Author information

1
UMR de Génétique Végétale, INRA/UPS/CNRS/INA-PG, Ferme de Moulon, F-91190 Gif-sur-Yvette, France.

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.

PMID:
15221754
DOI:
10.1002/pmic.200300731
[Indexed for MEDLINE]

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