Technique / Proteomics and protein biochemistry / Immunoprecipitation


Anti-FLAG Co-Immunoprecipitation



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)

Last update 17-Mar-2005, Rating of 5 votes.

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By Karen on 19-Sep-2008
I cannot help but it is comforting to know that I'm not the only one having this problem. Since February were you able to find a way to make the IP work?
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By Vikas on 15-Aug-2008
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
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By Deansy on 28-May-2008
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
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By Corinne Quittau on 21-Feb-2007
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?
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By Guo Xiangyun on 17-Mar-2005
this is a great lab helper to people , esp to the freshmen like me. thanks a lot！
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