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

被引：33

作者：

Fiévet, J

Dillmann, C

Lagniel, G

Davanture, M

Negroni, L

Labarre, J

de Vienne, D ^{[1
]}

机构：

[1] CNRS, INA PG, INRA, UMR Genet Vegetale, F-91190 Gif Sur Yvette, France

[2] CEA Saclay, Serv Biochim & Genet Mol, Gif Sur Yvette, France

[3] ISV, La Plante & Son Environm, IFR 87, Gif Sur Yvette, France

来源：

PROTEOMICS | 2004年 / 4卷 / 07期

关键词：

colloidal coomassie blue; image analysis; quantitative proteomics; two-dimensional gel electrophoresis; yeast;

D O I：

10.1002/pmic.200300731

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We statistically analysed various factors to get accurate estimates of protein quantities from two-dimensional gels. Yeast proteins were labelled with S-35 or stained with Coomassie Brilliant Blue G-250, and spots were automatically quantified with software packages Kepler, Image-QuaNT, Melanie 3.0 and Progenesis. The different software packages proved to have very similar performances. With S-35-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.

引用

页码：1939 / 1949

页数：11