Description
Principle of the Assay: This is an ELISA (Enzyme-Linked Immunosorbent Assay) for the quantitative analysis of testosterone levels in biological fluid. This test kit operates on the basis of competition between the enzyme conjugate and the testosterone in the sample for a limited number of binding sites on the antibody coated plate. The sample or standard solution is first added to the microplate. Next, the diluted enzyme conjugate is added and the mixture is shaken and incubated at room temperature for one hour. During the incubation, competition for binding sites is taking place. The plate is then washed, removing all the unbound material. The bound enzyme conjugate is detected by the addition of the substrate that generates an optimal color after 30 minutes. Quantitative test results may be obtained by measuring and comparing the absorbance reading of the sample wells against the wells of the standard curve with a microplate reader at 650 nm. The extent of color development is inversely proportional to the amount of testosterone in the sample or standard. For example, the absence of testosterone in the sample will result in a bright blue color, whereas the presence of testosterone will result in decreased or no color development.
Background/Introduction: Testosterone is the principle androgen. It is synthesized in the testis, the ovary and the adrenal cortex. It is responsible for the development and maintenance of the male secondary sex characteristics. It also exerts important protein anabolic and growth-promoting effects. The plasma testosterone levels are useful in investigating hypogonadism and hormone replacement therapy in men. It is also useful as a marker in hyper-androgenism in women