Neogen ELISA Drug Detection KitTestosterone 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.

This is an ELISA (Enzyme-Linked ImmunoSorbent Assay) for the quantitative analysis of Testosterone levels in biological fluid. This test kit operates based on 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 substrate which generates an optimal color after 30 minutes. Quantitative test results may be obtained by measuring and comparing the absorbance reading of the wells of the samples against the standards 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.

Standard Curve in EIA Buffer

Curve in EIA Buffer

Specificity (Cross-Reactivity)

Analytes Cross-Reactivity
Testosterone 100%
Dihydrotestosterone 100%
Androstenedione 0.86%
Bolandiol 0.86%
Testosterone Enanthate 0.13%
Estriol 0.10%
Testosterone Benzoate 0.10%
Estradiol 0.05%
Dehydroepiandrosterone 0.04%
Testosterone Propionate 0.04%
Deoxycorticosterone 0.03%
Testosterone 17β-Cypionate 0.02%
Aldosterone < 0.01%
Corticosterone < 0.01%
Cortisol < 0.01%
Cortisone < 0.01%
Estrone < 0.01%
17-Hydroxyprogesterone < 0.01%
Pregnenolone < 0.01%
Progesterone < 0.01%


   80% B/B0 = 0.006 ng/mL
   50% B/B0 = 0.029 ng/mL
Assay Range: 0.002 - 0.2 ng/mL
Inter-Assay CV: ≤ 10%
Intra-Assay CV: ≤ 10%
Size: 96 well microplate
Assay Sample Size: 50 µL
Total Assay Incubation Time: 1.5 hours
Wavelength: 650 nm, 450 nm with acid stop
Storage Conditions: 4°C
Antibody: Polyclonal, Rabbit

Order Details

Product # Product Description
402510 Testosterone Kit, 96 well kit

Reference Materials

402510 - Testosterone, Manual de instruções [Inglês]

Kit inserts available on our website are for reference use only and are subject to change. It is always recommended to use the protocol included with the kit.

Calculation of Results

Neogen has partnered with to provide an online tool for data calculations.

Follow the simple step-by-step guide to generate your data.

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Calculate Results is a service of MyAssays Ltd. For questions or concerns about the service, please contact MyAssays Ltd.

For research use only.

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