Free Testosterone

The Free Testosterone ELISA is an enzyme immunoassay for the quantitative in vitro measurement of Free Testosterone in serum and plasma.


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Product Catalog No: EIA-2924 Pack Size: 96 Wells

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Summary

Testosterone is a steroid hormone from the androgen group. Testosterone is primarily secreted in the testes of males and the ovaries of females although small amounts are secreted by the adrenal glands. It is the principal male sex hormone and an anabolic steroid. In both males and females, it plays key roles in health and well-being. Only 1-2% of circulating testosterone exists as unbound or free testosterone. The majority, approximately 60%, is bound to SHBG with high affinity, while the remainder is loosely bound to albumin. Both the albumin-bound and free fractions may be biologically active, while SHBG effectively inhibits testosterone action. Testosterone effects can be classified as virilizing and anabolic effects. Anabolic effects include growth of muscle mass and strength, increased bone density and strength, and stimulation of linear growth and bone maturation. Virilizing effects include maturation of the sex organs. Testosterone levels decline gradually with age in men. Measurement of the free or unbound fraction of serum testosterone has been proposed as a means of estimating the physiologically bioactive hormone. Free testosterone levels are elevated in women with hyperandrogenism associated with hirsutism in the presence or absence of polycystic ovarian disease. In addition, free testosterone measurements may be more useful than total testosterone in situations where SHBG is increased or decreased (e.g. hypothyroidism and obesity).

Test Principle

The Free Testosterone ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA), based on the principle of competitive binding. The microtiter wells are coated with an antibody directed towards an antigenic site on the Testosterone molecule. Endogenous Free Testosterone of a patient sample competes with a Testosterone-horseradish peroxidase conjugate for binding to the coated antibody. After incubation the unbound conjugate is washed off. The amount of bound peroxidase conjugate is inversely proportional to the concentration of Free Testosterone in the sample. After addition of the substrate solution, the intensity of colour developed is inversely proportional to the concentration of Free Testosterone in the patient sample. Testosterone in the blood is bound to SHBG (60 %) and in lower quantity to other protein. Only the measurement of Free Testosterone (< 1% of Total Testosterone) permits the estimating of the hormone biologically active.

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    References
    • McCann D, Kirkish L. J. Clin. Immunoassay 8:234-6 (1985)
    • Ekins RP., J. Clin. Immunoassay 1984; 7(2): 163 – 80
    • Paulson JD, et al., Am. J Obst. Gynecol 1977;128:851-7
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