Free Testosterone ELISA for serum or plasma 0.06-100.0 pg/ml Dialab

Testosterone is a steroid hormone form the androgen group secreted by the testes, ovaries, and in small amounts 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. Due to its insolubility in aqueous solutions, for the most part testosterone circulates in the blood bound to transport proteins. Only a small percentage (<1%) of circulating testosterone exists as unbound or free testosterone. The majority (~60%) is bound to sex hormone-binding globulin (SHBG) with high affinity; the remainder is loosely bound to albumin. Both the albumin-bound and free fractions may be biologically active, while SHBG effectively inhibits testosterone action. 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 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).
Normal male 4.2-42 pg/ml
Female:
Ovulating: ND – 4.1 pg/ml
Oral Contraceptives 0.3-2.0 pg/ml
Post meniopausal 0.1-1.7 pg/ml
This assay is intended for the quantitative determination of free testosterone in human serum or plasma for diagnostic use. This 96 well ELISA can provide up to 40 patient tests. Free testosterone in the sample competes with horseradish peroxidase (HRP) labelled testosterone for binding onto the limited number of anti- testosterone sites coated on the microplate. After incubation, the bound/free separation is performed by a simple- solid-phase washing. The enzyme in the bound-fraction reacts with the substrate to develop a blue colour which changes to yellow on addition of the stop solution. The colour intensity, as determined by reading absorbance at 450nm, is inversely proportional to the concentration of free testosterone in the sample.
Antibody coated Microtiter wells (Anti- Testosterone IgG): 12x8 well strips
Calibrator set (0.0, 0.2, 1.0, 4.0, 20.0, 100.0 pg/ml x 1.0ml)
Control: 1x1ml
Enzyme Conjugate (Anti-Testosterone-HRP): 1x15ml
Substrate Solution (H₂O₂-TMB 0.26g/L): 1x15ml
Stop Solution (Sulphuric Acid 0.15mol/L): 1x15ml
Wash Buffer 10X(0.2M Phosphate buffer): 1x50ml
Sample volume: 20µl human serum or plasma to be run in duplicate
Incubation time: 75 minutes (60 min at room temperature, 15 min in the dark at room temperature)
Measuring range: 0.06 – 100.0 pg/ml
Sensitivity: 0.06 pg/ml
Specificity: Testosterone 100%, DHT 0.00008%, Androstenedione 0.0043%, Androsterone 0.00029%,17β Oestradiol 0.00005%, DHEA-S 0.00007%, Oestrone < 0.00001%, Cortisol < 0.00001.
Store at 2-8°C.