Chapter title |
Co-immunoprecipitation from Transfected Cells
|
---|---|
Chapter number | 25 |
Book title |
Protein-Protein Interactions
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2425-7_25 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2424-0, 978-1-4939-2425-7
|
Authors |
Yoshinori Takahashi, Takahashi, Yoshinori |
Abstract |
Co-immunoprecipitation (Co-IP) is one of the most widely used methods to identify novel proteins that associate with a protein of interest or to determine complex formation between known proteins. For this technique, a protein of interest is captured using a specific antibody. The antibody-bound protein, as well as any proteins bound to the protein of interest, is then precipitated using a resin (immunoprecipitation, IP). Proteins that are not bound to the protein of interest are then removed from the sample with a series of washes. The resulting immunocomplexes are then analyzed by immunoblot. As the requirements for protein-protein interactions vary, optimal experimental conditions for examining the interacting partners of different proteins of interest must be determined empirically. Once appropriate experimental conditions have been established, the IP/Co-IP procedure is simple and straightforward. In this chapter, a standard protocol for IP/co-IP, with several key factors for the success of IP/co-IP analyses, is discussed. |
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