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Differential gel exposure, a new methodology for the two‐dimensional comparison of protein samples
Author(s) -
MonribotEspagne Christelle,
Boucherie Hélian
Publication year - 2002
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/1615-9861(200203)2:3<229::aid-prot229>3.0.co;2-k
Subject(s) - proteome , chemistry , chromatography , silver stain , yeast , in vivo , staining , biochemistry , microbiology and biotechnology , biology , genetics
We describe a novel methodology for the comparison of protein samples called differential gel exposure (DifExpo). This method is based on the coelectrophoresis on a two‐dimensional (2‐D) gel of two protein samples. The samples are differentiated from each other by in vivo radiolabelling, using 14 C‐ and 3 H‐isotopes. After 2‐D separation and transfer on a polyvinylidene difluoride membrane, the 3 H/ 14 C ratio of each protein spot is determined by exposure to two types of imaging plates, one sensitive to 14 C and the other to both 14 C and 3 H. We showed that DifExpo allows us to compare the cellular levels of several hundred proteins of the yeast proteome. Its sensitivity is comparable to silver staining. We also showed that it can be used to investigate changes in the rate of synthesis of individual proteins.