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nonspecific control for immunoprecipitation
mitti
Posted 8/8/2009 2:26:27 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
Posted 8/9/2009 9:44:29 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.
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mitti
Posted 8/10/2009 12:55:17 PM
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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?
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Pippuri
Posted 8/10/2009 1:05:03 PM
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Yes, it is. You can purchase it from many companies that provide antibodies.
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mitti
Posted 8/11/2009 2:37:17 AM
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Thank you very much for your help.
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