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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
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.
Harlow, E., and Lane, D. (1999) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc.
About this protocol
Cite this protocol
Lee, C. (2007). Coimmunoprecipitation Assay. In: Rosato, E. (eds) Circadian Rhythms. Methods in Molecular Biology™, vol 362. Humana Press. https://doi.org/10.1007/978-1-59745-257-1_31
Download citation
DOI: https://doi.org/10.1007/978-1-59745-257-1_31
Publisher Name: Humana Press
Print ISBN: 978-1-58829-417-3
Online ISBN: 978-1-59745-257-1
eBook Packages: Springer Protocols