WB using Anti-Wnt1 antibody shows detection of endogenous Wnt1 in human-derived MCF7 cell lysate (lane 1) and mouse-derived 3T3 cell lysate (lane 2). The band at ~41 kDa, indicated by the arrowhead, corresponds to Wnt1. After transfer, the membrane was blocked with 5% BLOTTO. Primary antibody was used at a 1:1, 400 dilution in PBS containing 1% BLOTTO. The specificity of the antibody was confirmed by peptide competition which completely blocked reaction of the antibody with Wnt1 (data not shown).