But the protocol is first to bind antibody to the beads and then bind your antigen. What you described here is you mixed antigen and antibody first and then added beads.
I tried both ways, did not see a difference. What is the antibody and lysate ratio you usually use?
n*w
8 楼
okay. so in this case, maybe your HSP90 antibody or other antibodies don't work well in IP conditions? I have met multiple occasions where IP only goes one-way... :-/ It could just be antibody issue. You can try cross link. If that doesn't work, then move on...