Figure 1

Characterization of PRLR isoform antibodies. A. Western blot analysis indicating specificity of polyclonal antibodies. CHO cells were transiently transfected with isoform specific PRLR cDNA as described. Cell lysates were prepared and proteins separated by PAGE. Each isoform specific lysate was probed with each isoform specific antibody. Each transfection was performed in triplicate and Western blot analysis performed twice. Data shown are a representative autoradiogram. Molecular weights are marked as indicated: LF = 93 kDa, SF1a = 57 kDa, SF1b = 38 kDa. Cell lysates from CHO transfected cells were immunoprecipitated with their respective antibodies, then immunoblotted with the same antibodies. Each transfection was performed in triplicate and Western blot analysis performed twice. Data shown are a representative autoradiogram. IP indicates immunoprecipitation. B. Western blot analysis using commercially available PRLR antibody. ECD = antibody recognizing extracellular domain C. Fluorescent immunocytochemical analysis. CHO cells were transiently transfected with isoform specific PRLR cDNA as described. Specific staining was observed using isoform specific polyclonal antibodies. The negative control (not shown) lacks primary antibody and appears black. Data shown are representative of triplicate experiments. Magnification 20×.