Overview: Co-immunoprecipitation (Co-IP) is a popular technique for the analysis of protein interaction. The procedure includes: 1) An antibody specific to the protein of interest is added to a cell lysate. 2) The antibody-protein complex is then precipitated usually using protein-G or protein-A sepharose which binds most antibodies. If there are any protein/molecules that bind to the first protein, they will also be precipitated. 3) Co-precipitated protein can then be identified by Western blot analysis or by sequencing a purified protein band.
Protocols
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Anti-FLAG Co-Immunoprecipitation Protocol
(Lykke-Andersen Lab, Department of MCD Biology, University of Colorado)
Procedures include cell transfection, immunoprecipitation and Western blot analysis.
http://www.colorado.edu/mcdb/Lykke_Andersen/Protoc...
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Co-Immunoprecipitation
(Lazo Lab, University of Pittsburgh)
http://www.pitt.edu/~lazo/Co-Immunoprecipitation.h...
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Co-Immunoprecipitation Analysis of Protein-Protein Interaction
(Morrisey Lab, University of Pennsylvania)
This protocol uses total cell extracts to analyze putative protein-protein interactions in eukaryotic cells. One can also use nuclear extracts as a source for this protocol.
http://www.uphs.upenn.edu/mcrc/morrisey/coip
Added: Mon Oct 14 2002, Hits: 20613, Reviews: 0 Write review Cached