can antibodies recommended for western blot be used for immunopreipitation - (Mar/10/2014 )
hello everbody,
I am sudheer. I am working on a protein for which the antibodies are available for western blot and ELISA. I need to do immunopreciptiation and co- IP studies on my protein. I have searched in web for antibodies., i found the antibodies recommended for WB and ELISA but not for IP and co-IP. I request you to please provide me the information regarding whether the antibodies for WB can also be used for IP and Co-IP .
thankyou.
The answer to this is: maybe
For IP you need an antibody that detects the native form (folded) of the protein, which is not the form you have in western blots, these are denatured. ELISA is usually done with native protein, so those may work, but it all depends on the antibody. Your best bet is to get some samples and try it
Hello Sudheer,
I agree that the answer to your question is a maybe and trying the antibody out will be a helpful approach - but you should research the antibodies in question a little further to help select which ones to try. If the immunogen (antigen) used to make the antibody is a protein and NOT a peptide, you have a higher likelihood of obtaining an antibody that will function in IP. Further, if the antibody is demonstrated to work in IHC, ICC or IF then there is a higher likelihood that it could work in an IP/co-IP application, as it can then recogonize the non-denatured form of the protein.