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2013•Cold Spring Harbor ProtocolsRequires access

Electrophoretic Mobility-Shift Assays

Michael Fionn Carey, Craig L. Peterson, Stephen T. Smale

Open publisher page 51 citations

Abstract

In an electrophoretic mobility-shift assay (EMSA, or simply "gel shift"), a (32)P-labeled DNA fragment containing a specific DNA site is incubated with a cognate DNA-binding protein. The protein-DNA complexes are separated from free (unbound) DNA by electrophoresis through a nondenaturing polyacrylamide gel. The protein retards the mobility of the DNA fragments to which it binds. Thus, the free DNA will migrate faster than the DNA-protein complex. An image of the gel is used to reveal the positions of the free and bound radiolabeled DNAs.

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What this paper is about

In an electrophoretic mobility-shift assay (EMSA, or simply "gel shift"), a (32)P-labeled DNA fragment containing a specific DNA site is incubated with a cognate DNA-binding protein. The protein-DNA complexes are separated from free (unbound) DNA by electrophoresis through a nondenaturing polyacrylamide gel. The protein retards the mobility of the DNA fragments to which it binds. Thus, the free DNA will migrate faster than the DNA-protein complex. An image of the gel is used to reveal the positions of the free and bound radiolabeled DNAs.

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OpenAlex reports 51 citations for this work. Citation counts describe recorded attention and do not establish research quality.

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Available abstract

In an electrophoretic mobility-shift assay (EMSA, or simply "gel shift"), a (32)P-labeled DNA fragment containing a specific DNA site is incubated with a cognate DNA-binding protein. The protein-DNA complexes are separated from free (unbound) DNA by electrophoresis through a nondenaturing polyacrylamide gel. The protein retards the mobility of the DNA fragments to which it binds. Thus, the free DNA will migrate faster than the DNA-protein complex. An image of the gel is used to reveal the positions of the free and bound radiolabeled DNAs.

Key concepts: Electrophoretic mobility shift assay, DNA, Electrophoresis, Gel electrophoresis of nucleic acids, Chemistry, Gel electrophoresis, Molecular biology, Polyacrylamide gel electrophoresis

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