enriching non-specific binders - (Aug/28/2006 )
I am doing pahge display on a quite complex antigen. What I observe now is that I am not only enriching specific binders, but also non-specific ones. To avoid this i already switched the blocking solution between every panning round from 3%BSA to 5% skimmed milk in the 2nd round and back to 3%BSA in the third. What I got in the polyclonal phage ELISA are antigen binders, but also increasing BSA binders and skimmed milk binders.
Any ideas how I can avoid enriching this crap??????
I doing exactly what you're doing but I block my immunotubes with 2%skimmed milk powder for 1hour and when I do polyclonal ELISA I get very good results without any background. I've never used BSA to block the tube, I'll suggest you try 2% milk powder.