Anti SMAD1, phospho Ser206  

Western blot using Rockland's Affinity Purified anti-SMAD1 pS206 antibody shows detection of endogenous phosphorylated SMAD1 in whole cell lysates from human hepatoma (HEPG2, lanes 1-4) derived cell lines treated with PBS, BMP2 (5 ng/mL), EGF (1 ng/mL), or NaCl for 1 h at 37°C before harvest. Each lane contains approximately 15 μg of lysate. Primary antibody was used at a 1:500 dilution in 1% BLOTTO (p/n B501-0500) and reacted for 1 hour at room temperature. Primary antibody was pre-incubated before reacting with blot as follows: top row - with PBS, middle row - with the immunizing phosphorylated peptide and bottom row - with control or non-phosphorylated peptide. The membrane was washed and reacted with a 1:3,000 dilution HRP-conjugated a-Rabbit IgG (p/n 611-103-122) for 1 hour at room temperature. Detection was by ECL. Personal communication, Xin-Hua Feng, Baylor College of Medicine, Houston, TX.