protein complexes are immunoprecipitated without antibody contamination. Manufacturer: Thermo Scientific™ PI Catalog No. PI $ / Each. It is the Pierce kit.I have also read articles where the kit was used successfully, but it does not seem to work for me. I am loosing my. pierce pdf reader. Quote. Postby Just» Tue Aug 28, am. Looking for pierce pdf reader. Will be grateful for any help! Top.
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Can someone who has had success performing Co-IPs help me troubleshoot problems I am experiencing. I have been trying to carry out a Co-IP for several months now, but I believe that either my kit or my antibody is sub-optimal.
Can someone suggest a good Co-IP kit to use which has proven successful for them? If you really want a co-IP kit, there is a selection to choose from. I never brought a kit for co-IP.
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If you have done IP before, co-Ip is just to “tune-down” your detergents in the lysis and wash buffers, otherwise the steps are the same. Thank you for your response. I have actually used one the Co-IP kits on the list you sent the link to, but I am having problems with it. It is the Pierce kit. I have also read articles where the kit was used successfully, but it does not seem to work for me.
I am loosing my protein before elution, which means that there is a problem with the antibody binding to the resin I think?
I am following the protocol to the letter, but I am loosing my protein. I suspect it is the antibody.
How do you ensure that the antibody you use will be optimal for the reaction? Now, the real issue that you are encountering is- whether your antibody is suitable for IP?
Have you tried to address this issue before proceeding to the co-IP experiment? Yes, I have been doing the IP now with this antibody.
You are correct, I have a feeling it is the antibody as well, but I have already purchased a new antibody and made sure that it was purified and dissolved in PBS, not Tris. Those were the 2 main criteria for selecting an antibody.
That is why I am not sure why it is still not working. I do detect it, but in the 2nd wash which suggests that I have lost all my protein before elution. The buffers are all pre-optimized for this kit and should not be too stringent for the reaction.
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If it is the antibody, how do I go about ensuring that I have purchased the right antibody? I would appreciate any insight into this problem since I am able to figure out where the problem lies. Log in or register to post comments. Hi Sue, If you really want a co-IP kit, there is a selection to choose from. Yes, I have been doing the IP.