I need a procedure for immunoprecipitation, that will enable me to obtain the immunoprecipitated proteins in the native state, so that I can perform enzyme activities and several spectroscopies to the purified samples.

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Did you try the protocols page on this website? Also try "Current protocols in molecular biology" and "Nature Protocols".

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I tried on this site and all procedures are based on binding to protein A sepharose, followed by sds solubilization.

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it's a long procedure and I don't have it on my pc to paste for you, but just a short recommendations:

1- try to use antibodies raised in two different species. e.g. IP ab raised in mice, WB ab raised in rabbit, so that your secondary antibody won't recognize both (trust me on this one, I used same species and I ruined my IP's)

2- use protein A beads.

3- rotate your tubes end to end.

4- bind your antibody to your protein A before adding your lysates or your protein sample including your protein of interest.


it is a very long and difficult experiment so don't expect to get result in the first attempt.

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