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Thread: Co-immunoprecipitation

  1. #1 Co-immunoprecipitation 
    Forum Freshman
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    HI THERE

    I'll be doing a CO-IP for first time, a protein of 60 kDa and another of 53 kDa, usingprotein G, what negative control should I use? also do not understand when paper say "control IgG", What concerns it?

    help!!!


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  3. #2  
    Forum Bachelors Degree
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    Hi kitty,

    I have never done a co-ip before unfortunately. I think that IgG is used as a control to ensure that your antibody is specifically recognising an epitope on your protein of interest (that is, the antigen), rather than the antibody recognising an epitope that is common to several different proteins. I think.

    This thread controls for Immunoprecipitation - Forum Thread - Biocompare Buyer's Guide For Life Scientists may help.

    Best wishes,

    Tri


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  4. #3  
    Forum Bachelors Degree
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    Or rather, to ensure that the IgG does not recognise your protein of interest.

    Can anyone else help with this?

    Tri
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  5. #4  
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    The IgG control is another antibody that recognizes a different antigen than you target to make sure that whatever you pull down is not binding non-specifically to the part of the antibody that does not bind to its antigen specifically. That is, if you see a target co-IP in the IgG control then it is not specific, it is a protein that binds antibodies, regardless of the antigen.
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