Calbiotech Androstenedione Elisa Kit

INTENDED USE
The Androstenedione Elisa Kit is intended for the measurement of Androstenedione in serum or plasma.

PRINCIPLE OF THE TEST

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