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Updated: Apr 15, 2026
Ultra-High-Speed Western Blot using Immunoreaction Enhancing Technology
Published on: September 26, 2020
Studying protein-protein interactions via blot overlay/far western blot
1Department of Pharmacology, Emory University School of Medicine, Rollins Research Center, Room 5113, 1510 Clifton Road, Atlanta, 30322, GA, USA, rhall@pharm.emory.edu.
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.
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Related Concept Videos
01:15Western Blotting
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
02:04Protein-protein Interfaces
02:26Protein Networks
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...