conditioned media for western - (Jan/27/2006 )
Hi, i need help in this one
I am thinking of analyzing conditioned media as I believe that the protein that I am trying to find in whole cell extracts is acively secreted. I just found some protocols about precipitating my hormone with antibody and then with protein A agarose for a latter western
Could some one tell me if I am in the right track? and if so, do you think I can store the media until the analyiss at -80'?
thanks in advance
We used EtOH to precipitate proteins, were concentrating secretions from ALI cultures and from the unbound CHIP fractions, (works alot like DNA precipitation) worked well for both of these, I would also check this link from protocol online... Don't know about freezing I think it depends on your protien of interest....