Detection of protein-protein interactions by coprecipitation

Current Protocols in Neuroscience
Elaine A Elion

Abstract

Coprecipitation of proteins from whole-cell extracts is a valuable approach to testing for physical interactions between proteins of interest. When a precipitating antibody is used, this method is referred to as co-immunoprecipitation. Coprecipitation can be used to study interactions between known proteins and as a means of identifying components of a complex. This unit describes basic approaches to immunoprecipitating tagged proteins from whole-cell extracts. The extract is prepared under nondenaturing conditions, the protein of interest is precipitated, and the precipitate is tested for a second specifically associated protein. The approach can be used for native or epitope-tagged proteins for which antibodies are available, or for recombinant proteins that bind with high affinity to a molecule that can be coupled to a solid-phase matrix. An associated protein is detected by separating the precipitated proteins by SDS-PAGE and immunoblotting with a second antibody that recognizes the putative associated protein.

References

Jan 1, 1991·Methods in Enzymology·P A Kolodziej, R A Young
Oct 19, 2000·Methods in Enzymology·J A Bornhorst, J J Falke
Jun 14, 2001·Methods : a Companion to Methods in Enzymology·G G Toby, E A Golemis
Jun 14, 2001·Methods : a Companion to Methods in Enzymology·O PuigB Séraphin
Dec 31, 2002·Journal of Receptor and Signal Transduction Research·Daniel AuerbachIgor Stagljar
Jan 28, 2003·Current Opinion in Chemical Biology·Anne-Claude Gavin, Giulio Superti-Furga
Apr 23, 2008·Current Protocols in Protein Science·G Michaels, R Garian

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Citations

Dec 30, 2014·Mass Spectrometry Reviews·Elien VandermarliereLennart Martens
May 15, 2013·Acta Biochimica Et Biophysica Sinica·Ying ZhangYuan Lin

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