DHEA-S ELISA Kit
DHEA-S ELISA Kit
The DHEA-S ELISA kit is based on the principle of competitive binding between DHEA-S in the test specimen and HRP conjugated DHEA-S for a constant amount of anti-DHEA-S antibody. In the first incubation, Goat-anti-Rabbit-IgG coated wells are incubated with 10uL of DHEA-S standards, patient samples, 50uL DHEA-S Enzyme (HRP) reagent and 50uL anti-DHEA-S Antibody reagent, at room temperature, for 60 minutes. During the incubation, HRP labeled DHEA-S competes with the endogenous DHEA-S in the standard and sample, for a fixed number of binding sites of the DHEAS antibody, while simultaneously the Anti DHEA-S antibody binds to the immobilized secondary antibody. Thus, the amount of DHEA-S HRP conjugate immunologically bound to the well progressively decreases as the concentration of DHEA-S in the specimen increases. Unbound DHEA-S HRP conjugate is then removed and the wells washed. Next, TMB Reagent is added and incubated at room temperature for 30 minutes, resulting in the development of blue color. The color development is stopped with the addition of stop solution, and the absorbance is spectrophotometrically measured at 450nm. A standard curve is prepared relating color intensity to the concentration of DHEA-S.
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