Free Testosterone ELISA Kit

Free Testosterone ELISA KitThe Free Testosterone ELISA kit is based on the principle of competitive binding between testosterone in the test specimen and Testosterone HRP conjugate for a constant amount of rabbit anti Free Testosterone. In the incubation, goat anti rabbit IgG coated wells are incubated with 25uL of Testosterone standards, patient samples, 50uL testosterone HRP conjugate reagent and 50uL rabbit anti free testosterone reagent at room temperature for 60 minutes.

The Free Testosterone ELISA kit is based on the principle of competitive binding between testosterone in the test specimen and Testosterone-HRP conjugate for a constant amount of rabbit anti-Free Testosterone. In the incubation, goat anti-rabbit IgG-coated wells are incubated with 25uL of Testosterone standards, patient samples, 50uL testosterone-HRP conjugate reagent and 50uL rabbit anti-free testosterone reagent at room temperature for 60 minutes. During the incubation, a fixed amount of HRP labeled testosterone competes with the endogenous testosterone in the standard and sample, for a fixed number of binding sites of the specific free testosterone antibody. Thus, the amount of testosterone peroxidase conjugate immunologically bound to the well progressively decreases as the concentration of free testosterone in the specimen increases. Unbound testosterone peroxidase conjugate is then removed and the wells washed. Next, 100uL of TMB Reagent is added and incubated at room temperature for 15 minutes, resulting in the development of blue color. The color development is stopped with the addition of 50uL stop solution, and the absorbance is measured spectrophotometrically at 450nm. A standard curve is prepared relating color intensity to the concentration of the Free Testosterone.