Neogen ELISA Histamine Kit17β-Estradiol is considered the true ovarian hormone in women. It is the most potent estrogen, including Estrone and Estriol. 17β-Estradiol is responsible for the reproductive epithelia, breast growth, maturation of long bones, and development of the secondary female sex characteristics. The 17β-Estradiol level in plasma is used in assaying for fertility, amenorrhea and precocious puberty in girls.

This is an ELISA (Enzyme-Linked ImmunoSorbent Assay) for the quantitative analysis of Estradiol levels in biological fluid. This test kit operates based on competition between the enzyme conjugate and the Estradiol 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 Estradiol in the sample or standard. For example, the absence of Estradiol in the sample will result in a bright blue color, whereas the presence of Estradiol will result in decreased or no color development.

Standard Curve in EIA Buffer

Curve in EIA Buffer

Specificity (Cross-Reactivity)

Analytes Cross-Reactivity
17β-Estradiol 100%
Testosterone 1%
Estriol 0.41%
Estrone 0.10%
Dehydroepiandrosterone 0.03%
Aldosterone < 0.02%
Androstenedione < 0.02%
Corticosterone < 0.02%
Cortisol < 0.02%
Cortisone < 0.02%
Deoxycorticosterone < 0.02%
17-Hydroxyprogesterone < 0.02%
Pregnenolone < 0.02%
Progesterone < 0.02%
17α-Ethynylestradiol < 0.01%
β-Estradiol 17-(β-D-glucuronide) < 0.01%


   80% B/B0 = 0.03 ng/mL
   50% B/B0 = 0.28 ng/mL
Assay Range: 0.02 - 2.0 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
402110 Estradiol Kit, 96 well kit

Reference Materials

402110 - Estradiol, Instrucciones del kit [Inglés]

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

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