I am trying to do immuno-precipitations in denaturing conditions. One protocol I have says denature in a buffer with 1.85 M NaOH + 7% β-Mercaptoethanol, then precipitated with 50 %TCA. Again the pellet was denatured with buffer containing 2% SDS. Next dilute the lysate to 1:5 and IP using anti HAaffinity matrix (my protein is tagged with HA). But I don’t have anti-HA affinity matrix so I want to do IP using anti HA antibody. Any one tried IP using antibodies in denaturing conditions? The antibodies will be stable in these conditions?Let me know if it works
Thanks