Principle of Assay
Human Denosumab ELISA Kit (A303039) employs the indirect sandwich enzyme immunoassay technique for the quantitative measurement of human Denosumab in serum, plasma, tissue homogenates, and other biological fluids. A capture antibody specific for Denosumab is pre-coated onto the 96-well microtiter plate. Samples and standards are added, and Denosumab present in each sample binds to the immobilized antibody. After washing, a Biotinylated detection antibody is added and incubated. Following washing, HRP-Streptavidin is added and incubated. TMB substrate is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of Denosumab captured, and the concentration is calculated by reading O.D. at 450nm and referring to the standard curve.