Crosslinking Ab w/Antigen? - (Sep/02/2009 )
I'm seeking some advice: I would like to permanently crosslink a monoclonal Ab I have against a known antigen. Keep in mind that this mAb will be biotinylated.
Here is the thing...would I be wise in just performing a quick UV crosslinking step in solution to help ensure they are permanently attached? The UV should not harm my biotin tag. For my experiment, I want to ensure the antigen:mAb remains permanently stable and I would like to avoid any unnecessary structural modifications. I want both the antibody and antigen to be in their most native states. Any insight would be greatly appreciated.
Any thoughts and/or suggestions would be greatly appreciated.
Did you mean that you will use Ag labeled with photoactivatable cross-linker plus UV treatment (prefered) or just UV? Pierce carries all kind of cross-linkers.
genehunter on Sep 2 2009, 08:58 AM said:
At this point, I was considering just UV as I don't have the Ag labeled with any photoactivatable cross-linkers. I don't have tons of the mAb (as always tends to be the case), but what I do have is already biotinylated. At the least, the UV treatment shouldn't alter that.