Calbiotech 17(OH) Progesterone ELISA

PRINCIPLE OF THE TEST 

The Calbiotech 17α-OH Progesterone (17-OHP) is a based on the principle of competitive binding between 17- OHP in the test specimen and 17-OHP-HRP for a constant amount of Rabbit anti-17-OHP antibody. In the assay, goat anti-rabbit IgG-coated wells are incubated with 25µl 17-OHP standards, 50µl 17-OHP-HRP conjugate, and 50µl rabbit anti-17-OHP. HRP-labeled-17-OHP competes with 17-OHP in the standards and sample for a fixed number of binding sites of the specific anti-17-OHP antibody. Thus, the amount of 17-OHP peroxidase conjugate immunologically bound to the well progressively decreases as the concentration of 17- OHP in the specimen increases. Unbound 17-OHP is then removed by the washing steps. TMB Substrate is added, 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 17-OHP.