Anti-FLAG Co-Immunoprecipitation
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Anti-FLAG Co-Immunoprecipitation
Immunoprecipitation is a technique that permits the purification of specific proteins for which a antibody has been raised. This primary antibody is either already bound to agarose or can be bound to the protein A/agarose beads during the procedure in order to physically separate the antibody-antigen complex from the remaining sample. (Laboratory of Gene Therapy Research, Copenhagen University Hospital)
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Last update 17-Mar-2005, Rating Good of 4 votes.
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Dear Deansy,
Thanks for your answer. Could you explain at what step in IP Arginine must be added, and if possible, how it works in the solution?
Thanks
Rating: Very Good
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Add 100mM Arginine to your IPs. Non-specific background is mostly caused by coprecipitation of aggregated proteins, and Arginine will reduce aggregation without interfering with specific protein:protein interactions
Rating: Good
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We used the anti-flag M2 affinity gel from sigma in IP experiments of cellular extracts from cells transfected with a flagged protein. Bad surprise: IPs revealed by western blot with an anti-flag antibody showed multiple bands , suggesting that the anti-flag M2 affinity gel immunoprecipate proteins able to interact with the anti-flag antibody. By contrast the supernatent did not exhibit any bands recognized by the anti-flag antibody. Can anybody help us?
Rating: Poor
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this is a great lab helper to people , esp to the freshmen like me. thanks a lot！
Rating: Excellent!
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