A recombinant variant of the human TSH receptor (70.8 kDa) was subjected to Western blotting. Upon incubation with the primary antibody (mixture of human Graves disease autoimmune sera), the membrane was washed and incubated with Goat anti-Human-AP (1:1,000). Colorimetric detection
Determination of human autoantibodies with Goat anti-Human-AP. A recombinant variant of the human TSH receptor was immobilized onto an ELISA-plate (coat, Co) and incubated with a mixture of human Graves disease autoimmune sera. Bound autoantibodies were determined with Goat anti-Human-AP at the indicated dilution factors. Visualization was performed with a commercially available PNPP ELISA substrate.