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Difference gel electrophoresis. A single gel method for detecting changes in protein extracts
Author(s) -
Ünlü Mustafa,
Morgan Mary E.,
Minden Jonathan S.
Publication year - 1997
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150181133
Subject(s) - difference gel electrophoresis , gel electrophoresis , polyacrylamide gel electrophoresis , chromatography , fluorescence , chemistry , electrophoresis , two dimensional gel electrophoresis , color marker , gel electrophoresis of proteins , molecular weight size marker , gel electrophoresis of nucleic acids , significant difference , microbiology and biotechnology , biology , biochemistry , proteomics , gene , physics , statistics , mathematics , quantum mechanics , enzyme
Abstract We describe a modification of two‐dimensional (2‐D) polyacrylamide gel electrophoresis that requires only a single gel to reproducibly detect differences between two protein samples. This was accomplished by fluorescently tagging the two samples with two different dyes, running them on the same 2‐D gel, post‐run fluorescence imaging of the gel into two images, and superimposing the images. The amine reactive dyes were designed to insure that proteins common to both samples have the same relative mobility regardless of the dye used to tag them. Thus, this technique, called difference gel electrophoresis (DIGE), circumvents the need to compare several 2‐D gels. DIGE is reproducible, sensitive, and can detect an exogenous difference between two Drosophila embryo extracts at nanogram levels. Moreover, an inducible protein from E. coli was detected after 15 min of induction and identified using DIGE preparatively.