I'm planning to do an IP experiment using the same antibody for both the IP and the western. I'm thinking of crosslinking the antibody to protein A beads using DMP. I'm wondering, apart from improving the resolution of the blot by keeping the light and heavy chains on the beads, is there any other reason to do the crosslinking? Does it improve the efficiency of the IP, and would people recommend it even if interference with light and heavy chains were not an issue?


Thanks for your input.

smu

as you said, crosslinking the ab to protein a will prevent the ab from eluting off the protein a when you elute the antigen from the ab. if you use a gentle method to elute then you may be able to reuse the beads.

if the presence of the binding ab won't interfere with your blot then you don't need to crosslink. but, if it will interfere then it is a good idea to crosslink.

i crosslink for immunoaffinity chromatography.