Issue 5, 2012

Identification of DNA–protein complexes using an improved, combined western blotting-electrophoretic mobility shift assay (WEMSA) with a fluorescence imaging system

Abstract

Electrophoretic Mobility Shift Assays (EMSAs) are used to detect DNA–protein interactions. With this type of assay it is difficult to distinguish between specific and non-specific DNA–protein complexes or to define which protein binds to the DNA. Here we describe a novel Western blot-combined EMSA (WEMSA) variant for a fluorescence imaging system which permits easy identification of specific DNA–protein-complexes. This method also allows investigation of several DNA–protein complexes in parallel. We have identified and distinguished clearly between the SP1- and EGR1-DNA–protein complexes which exhibit overlapping binding to the GC-BOX of the mPGES-1 promoter.

Graphical abstract: Identification of DNA–protein complexes using an improved, combined western blotting-electrophoretic mobility shift assay (WEMSA) with a fluorescence imaging system

Supplementary files

Article information

Article type
Method
Submitted
15 Dec 2011
Accepted
01 Feb 2012
First published
03 Feb 2012

Mol. BioSyst., 2012,8, 1389-1395

Identification of DNA–protein complexes using an improved, combined western blotting-electrophoretic mobility shift assay (WEMSA) with a fluorescence imaging system

K. Deckmann, F. Rörsch, G. Geisslinger and S. Grösch, Mol. BioSyst., 2012, 8, 1389 DOI: 10.1039/C2MB05500G

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