Application Notes: This Rat Vitamin D3 (VD3) ELISA Kit employs a two-site sandwich ELISA to quantitate VD3 in samples. An antibody specific for VD3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyVD3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for VD3 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of VD3 bound in the initial step. The color development is stopped and the intensity of the color is measured
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