title: Electrophoretic Mobility Shift Assay Using Radiolabeled DNA Probes.
abstract: Electrophoretic mobility shift assays (EMSA) have proven their usefulness for studying interactions between biological molecules. In the present protocol, a purified protein of interest is mixed with a 5'-end radiolabeled DNA probe. The bound complexes are separated by electrophoretic migration through a polyacrylamide gel and detected with a phosphorimager. The applications of EMSA are diverse, from thermodynamic and kinetic analyses to observation of bending and other conformational changes, stoichiometric inferences, or insights into cooperative protein binding.
authors.affiliation: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada, J1K 2R1.
authors.forename: Dominic
authors.initials: D
authors.lastname: Poulin-Laprade
authors.affiliation: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, QC, Canada, J1K 2R1. Vincent.Burrus@USherbrooke.ca.
authors.forename: Vincent
authors.initials: V
authors.lastname: Burrus
mesh_terms: D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D011485:Protein Binding; D011868:Radioisotopes
chemical_list: D015342:DNA Probes; D004268:DNA-Binding Proteins; D011868:Radioisotopes
keywords: DNA probe labeling; EMSA; Electrophoretic mobility shift assay; Gel retardation; Gel shift; Native polyacrylamide gel; Phosphorus-32; Protein–DNA interactions