co-immunoprecipitation of endogenous proteins

1 post / 0 new
k-lo's picture
co-immunoprecipitation of endogenous proteins

Hello everyone,
I am somewhat new to co-immunoprecipitation/western blotting and am mainly interested in co-immunoprecipitation of cell surface receptors. I would like to know if anyone has a standard protocol for co-immunoprecipitation of endogenous cellular proteins? I would also like to hear your opinions or suggestions on:

1. The amount of cells needed for a single IP (i.e. # of dishes, flasks etc).
2. concentration of antibody used for the IP.
3. The amount of Protein A/G agarose/sepharose used.
4. Elution volume and amount of material loaded in each lane.

Thanks for your time and I look forward to hearing from you.