heavy background in IP - (Nov/24/2010 )

Hi all:

I have been stuck in the immunoprecipitation for a long time. The protocol right now works, but there is a heavy background in the gel, which is extreme difficult for me to identify the associated protein. I plan to use DMP to crosslink the antibody to protein A beads. Does it work? Anyone has any experiences and suggestions? Thank you.

Posting your protocol should help diagnose any potential problems. Have you tried protein G beads?

we use a kit that uses dss to crosslink antibodies to protein a agarose (igg plus orientation kits).