Description
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Vitamin D3 (VD3) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Vitamin D3 (VD3) and unlabeled Vitamin D3 (VD3) (Standards or samples) with the pre-coated antibody specific to Vitamin D3 (VD3). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Vitamin D3 (VD3) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Vitamin D3 (VD3) in the sample