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nonspecific control for immunoprecipitation

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mitti
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Topic Started by mitti
on 8/8/2009 12:26 PM   
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Hi all,

I'm going to do immunoprecipitation on human ovarian cell line by a rabbit polyclonal antibody. I have been told to use an irrelevant polyclonal antibody as a nonspecific control. It is written in protocols to use an antibody to an epitope that is not present in the cell lysate as a control but I don't know what to use and how. does any body have experience in IP, what controls do you include in you IP?


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Pippuri
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Posted By Pippuri
on 8/9/2009 7:44 AM   
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 Mitti,

The most common negative control is to use normal rabbit IgG for IP in parallel, and compare to your experimental groups, normal rabbit IgG shouldn't pull down what you are looking for.

Good luck.



mitti
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Posted By mitti
on 8/10/2009 10:55 AM   
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Does normal rabbit IgG mean IgG from a rabbit that is not immunized? so it is not against any human epitope?



Pippuri
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Posted By Pippuri
on 8/10/2009 11:05 AM   
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Yes, it is. You can purchase it from many companies that provide antibodies.



mitti
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Posted By mitti
on 8/11/2009 0:37 AM   
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Thank you very much for your help.



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