The Testosterone ELISA measures Testosterone in samples. Add samples to the pre-coated plate. Then add biotinylated antigen. The antigens in the samples compete with the biotinylated antigen to bind to the capture antibody and incubate. Unbound antigen is washed away during a washing step. An avidin-HRP is then added and then incubate. Unbound avidin-HRP is washed away during a washing step. TMB Substrate is then added and color develops. The reaction is stopped by addition of acidic stop solution and color changes into yellow that can be measured at 450 nm. The intensity of the color developed is inversely proportional to the concentration of Testosterone in the sample. The concentration of Testosterone in the sample is then determined by comparing the O.D. of the samples to the standard curve.
Research AreasSignal Transduction, Endocrinology, Hormone Metabolism
BackgroundTestosterone is the primary sex hormone and anabolic steroid in males. In male humans, testosterone plays a key role in the development of male reproductive tissues such as testes and prostate, as well as promoting secondary sexual characteristics such as increased muscle and bone mass, and the growth of body hair. In addition, testosterone in both sexes is involved in health and well-being[5] and in the prevention of osteoporosis. Insufficient levels of testosterone in men may lead to abnormalities including frailty and bone loss. Testosterone is a steroid from the androstane class containing a keto and hydroxyl groups at positions three and seventeen respectively. It is biosynthesized in several steps from cholesterol and is converted in the liver to inactive metabolites. It exerts its action through binding to and activation of the androgen receptor. In humans and most other vertebrates, testosterone is secreted primarily by the testicles of males and, to a lesser extent, the ovaries of females. On average, in adult males, levels of testosterone are about seven to eight times as great as in adult females. As the metabolism of testosterone in males is more pronounced, the daily production is about 20 times greater in men. Females are also more sensitive to the hormone. Source: UniProt Consortium (2025)
