Studying protein-protein interactions via blot overlay or Far Western blot.
Blot overlay is a useful method for studying protein-protein interactions. This technique involves fractionating proteins on SDS-PAGE, blotting to nitrocellulose or PVDF membrane, and then incubating with a probe of interest. The probe is typically a protein that is radiolabeled, biotinylated, or simply visualized with a specific antibody. When the probe is visualized via antibody detection, this technique is often referred to as "Far Western blot." Many different kinds of protein-protein interactions can be studied via blot overlay, and the method is applicable to screens for unknown protein-protein interactions as well as to the detailed characterization of known interactions.
Hall
Randy A
RA
Department of Pharmacology, Emory University, Atlanta, GA, USA.
D000818:Animals; D026481:Blotting, Far-Western / Q000379:methods*; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D015335:Molecular Probes; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
D015335:Molecular Probes; D011506:Proteins