Figure 1. Specificity of the Ang II receptor antibodies.

Electrophoresis of renal membranes was carried out as described in the Materials and Methods section, using 80 µg of total protein. The specificity of the antibodies was confirmed by a preadsorption experiment using full-length human Ang II type 2 receptor recombinant protein as a matrix. The samples were incubated for 72 h at 4°C with gentle stirring and then centrifuged at 18,000×g for 1 min. The secondary antibody was a polyclonal anti-rabbit (NIF824, GE; 1∶2,500). See additional details in the Materials and Methods section. A: Ponceau red-stained nitrocellulose membrane (left) and immunosignal obtained after incubation of the AT₁R antibody with the full-length AT₂R recombinant protein (right). B: Ponceau red-stained nitrocellulose membrane (left) with no signal when the AT₂R antibody was preadsorbed on to the full-length AT₂R recombinant protein (right).