AMH, Bovine ELISA

The Bovine AMH ELISA is a quantitative three-step sandwich type immunoassay.  In the first step serially diluted Calibrators and unknown samples are added to AMH antibody coated micro titer wells and incubated. After the first incubation and washing, the wells are incubated with biotinylated AMH antibody solution. After second incubation and washing, the wells are incubated with streptavidin horseradish peroxidase conjugate (SHRP) solution.  After the third incubation and washing step, the wells are incubated with substrate solution (TMB) followed by an acidic stopping solution. In principle, the antibody-biotin conjugate binds to the solid phase antibody-antigen complex which in turn binds to the streptavidin-enzyme conjugate. The antibody-antigen-biotin conjugate-SHRP complex bound to the well is detected by enzyme-substrate reaction. The degree of enzymatic turnover of the substrate is determined by dual wavelength absorbance measurement at 450 nm as primary test filter and 630 nm as reference filter. The absorbance measured is directly proportional to the concentration of AMH in the samples and calibrators.