Anti-p27 Antibody - Western Blot. Western blot of affinity purified anti-p27 antibody shows detection of p27 protein in MCF7 whole cell lysate (lanes 1-3) (arrowhead). Separation was achieved using a 4-20% gradient gel. Blocking occurred using 5% BLOTTO. Primary antibody was diluted 1:500 in 1% BLOTTO. The membrane was washed and reacted with a 1:10000 dilution of Dylight 800 conjugated Gt-a-Rabbit IgG. Molecular weight estimation was made by comparison to prestained MW markers indicated at the left (lane M). Other detection systems will yield similar results.