Anti-SMAD1 Antibody - Western Blot. Western blot of Affinity Purified anti-SMAD1 antibody shows detection of endogenous SMAD1 in whole cell lysates from human hepatoma (HEPG2, lanes 1-4) and keratinocyte (HaCaT, lanes 5-8) derived cell lines treated with PBS, BMP2, EGF, or NaCl for 1 h at 37?C before harvest. Each lane contains approximately 15 ug of lysate. Primary antibody was used at a 1:500 dilution in 1% BLOTTO (B501-0500) and reacted for 1 hour at room temperature. Anti-beta actin (p/n LS-B323) staining was used as a loading control. The membrane was washed and reacted with a 1:3000 dilution HRP-conjugated a-Rabbit IgG (p/n LS-C60865) for 1 hour at room temperature. Detection was by ECL. Personal communication, Xin-Hua Feng, Baylor College of Medicine, Houston, TX.