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Coimmunoprecipitation Assay

  • Choogon Lee
Part of the Methods in Molecular Biology™ book series (MIMB, volume 362)

Abstract

As with most other proteins, clock proteins physically interact with one another. Coimmunoprecipitation (coIP) is the most straightforward technique to study protein-protein interactions in vivo, if antibodies against the proteins of interest are available. To perform coIP, first an antibody against a target protein is coupled to Sepharose beads through protein A or G, then the complexes containing the target protein are immunoprecipitated with the antibody-coupled beads by centrifugation. Protein components in the complexes are visualized by Western blotting using antibodies specific to the different components.

Key Words

Coimmunoprecipitation protein-protein interactions protein A/G protein complexes 

References

  1. 1.
    Kessler, S. W. (1975) Rapid isolation of antigens from cells with a staphylococcal protein A-antibody absorbent: Parameters of the interaction of antibody-antigen complexes with protein A. J. Immunol. 115, 1617–1624.PubMedGoogle Scholar
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    Akerstrom, B., Brodin, T., Reis, K., and Bjorck, L. (1985) Protein G: A powerful tool for binding and detection of monoclonal and polyclonal antibodies. J. Immunol. 135, 2589–2592.PubMedGoogle Scholar
  3. 3.
    Harlow, E., and Lane, D. (1999) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Choogon Lee
    • 1
  1. 1.Department of Biomedical SciencesFlorida State University College of MedicineTallahassee

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