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 virilising 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. Virilising 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).

May be diagnosed by:

• Determination of specific antibodies based on the ELISA technique
• Determination of specific antibodies based on the RIA-technique

Free Testosterone ELISA:

The Free Testosterone ELISA is intended for competitive immunoenzymatic colorimetric method for quantitative determination of Free Testosterone in human serum or plasma.

Principle of the Assay:

Microtiter strip wells are precoated with anti-Testosterone antibodies (solid-phase). Testosterone in the sample competes with added horseradish peroxidase labelled Testosterone (enzyme-labelled antigen) for antibody binding. After incubation a bound/free separation is performed by solid-phase washing. The immune complex formed by enzyme-labelled antigen is visualized by adding Tetramethylbenzidine (TMB) substrate which gives a blue reaction product. The intensity of this product is inversely proportional to the amount of Testosterone in the sample. Sulphuric acid is added to stop the reaction. This produces a yellow endpoint colour. Absorption at 450 nm is read using an ELISA microwell plate reader.

Specific performance characteristics: 

Correlation with another commercially available free testosterone assay performed on 95 samples is r = 0.86

Order information: