How many PBMCs should I seed so that I can get enough protein? - (Jul/17/2013 )

I was wondering if anyone could tell me how many PBMCs I need to seed in a 100mm dish in order to get enough PBMCs for protein extraction? I have previously seeded 5-6x10⁶ cells in a 12-well plate, and this gives me enough RNA for RT-PCR experiments.

I'm actually planning to seed the PBMCs, allow 2 hours to pass until the monocytes adhere and then remove the non-adherent cells (as per the materials and methods section seen in the paper that I have attached).

Any help would be very much appreciated.

Many thanks.

What sort of protein extraction? Are you just looking to make a cell lysate and do something like western blotting, or are you looking to purify a particular protein or do immunoprecipitation/co-IP?