phospho-Smad2/Smad3 (Thr8) Polyclonal Antibody

Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103 and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Smad2\/Smad3 (Thr8) Polyclonal Antibody(bs-8853R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).