Antibody Specificity Validation for Receptor Studies

The specificity of the antibodies used for WB and immunolabeling studies has been established in previous studies: AT1 sc-31181;^{59 (link)} AT2 sc-9040.^{60 (link)} In addition, the specificity of the antibodies was confirmed in our laboratory by pre-adsorption with the corresponding synthetic peptide antigen. See ref. 61 (link) for details. We also used WB analysis of lysates from HEK293 cells transfected with AT1 or AT2 tagged to fusion tail DDK (TA50011 from Origene, Rockville, MD, USA; DDK tag: DYKDDDDK). Using HEK293 cells the specificity of the antibodies was confirmed by the presence of a predominant immunoreactive band in lysates from positively transfected cells and also by the absence of this band in negative controls, which consisted of lysates of cells transfected with empty vectors. See ref. 17 (link) for details.