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PMID: 15064457 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Review

Studying protein-protein interactions via blot overlay or Far Western blot.

Methods in molecular biology (Clifton, N.J.) ·Vol. 261 ·2004-00-00 ·Pages 167-74

Hall RA

Abstract

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.

MeSH Terms
Animals Blotting, Far-Western/methods Electrophoresis/methods Humans Molecular Probes Protein Binding Protein Interaction Mapping/methods Proteins/chemistry,metabolism
Chemicals
Molecular Probes Proteins
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Hall Randy A
Department of Pharmacology, Emory University, Atlanta, GA, USA.
Article Info
Journal
Methods in molecular biology (Clifton, N.J.)
Abbr.
Methods Mol Biol
ISSN
1064-3745
Published
2004-00-00
Pages
167-74
Language
English
Region
United States
NLM ID
9214969
Subset
IM
Grants
NIGMS NIH HHS · GM 60982 · United States
NHLBI NIH HHS · HL 64713 · United States
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