Estradiol ELISA Kit
Estradiol ELISA Kit
The Estradiol (E2) ELISA kit is based on the principle of delayed competitive binding assay between E2 in the test specimen and E2 enzyme conjugate for a constant amount of anti-Estradiol monoclonal antibody epitope (Biotin reagent). In the incubation, anti-E2 antibody biotin reagent, E2 standards, controls, and samples are incubated for 45 minutes at room temperature (RT), then E2 enzyme conjugate is added on the top of the reaction mixture and incubation continues for 45 minutes more. During the incubation, a fixed amount of HRP-labeled E2 competes with the endogenous E2 in the standard, sample, or quality control serum for a fixed number of binding sites of the specific E2 antibody. E2 peroxidase conjugate immunologically bound to the well progressively decreases as the concentration of E2 in the specimen increases. Unbound of anti-Estradiol Biotin Reagent and E2 peroxidase conjugate is then removed and the wells are washed. Next, a solution of TMB Reagent is added and incubated at room temperature for 20 minutes, resulting in the development of blue color. The color development is stopped with the addition of stop solution, and the absorbance is measured spectrophotometrically at 450 nm. A standard curve is obtained by plotting the concentration of the standard versus the absorbance
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