Estradiol, 17b (E2), ELISA Kit

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Catalog Number: 40-056-205004

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Product Name: Estradiol, 17b (E2)

Category: Fertility

Sample Type: Serum and plasma

Sensitivity: 5 pg/mL

Description: Enzyme immunoassay for the quantitative measurement of Estradiol in serum and plasma

# of Samples: 1X96 Assays

Intended Use: The GenWay E2-EASIA is a competitive binding immunoassay for the quantitative determination of estradiol in serum and plasma.

Introduction: A. Estradiol
17-beta-estradiol (E2) is a C-18 steroid hormone (molecular weight 272.4) produced mainly by the ovary and placenta, and in small amounts by adrenals and testes. Estradiol is in equilibrium with estrone, which can be converted to estriol by the liver and placenta.
B. Clinical applications of the measurement of E2 levels
Like LH, FSH and progesterone, measurement of estradiol concentration in serum, peritoneal fluid and follicular fluid is an essential biochemical tool for the investigation of fertility, tumor and sexual diseases and disorders of hypothalamic/pituitary/gonadal axis.
For example:
- to detect the follicular phase;
- to check the effectiveness of the induction of ovulation (with ultrasound) and the level of E2 in follicular fluid. It allows normal detection or dysfunctional ovulation induction (the empty follicle synfrome may reflect a dysfonctional ovulation induction);
- to diagnose the luteinized unruptured follicle (LUF) syndrome (by the estimation of 17 betaestradiol and progesterone levels in peritoneal fluid);
- to aid in the diagnosis of breast tumors (total estrogens – E1-E2 – and 17 beta-hydroxysteroïd dehydrogenase activity are significantly higher in malignant than in non malignant breast tissues);
- other areas of investigation are: premature adrenarche, gynecomastie and menopausal period.

Principle of the test: E2-EASIA is an enzyme immunoassay performed in a microtiter plate. A fixed amount of estradiol labeled with horseradish peroxidase (HRP) competes with unlabeled estradiol present in calibrators or samples for a limited number of binding sites of a specific antibody. The E2-HRP-antibody complex is simultaneously fixed on the wells of the microtiter plate coated with an excess of anti-rabbit-gammaglobulins.
Neither extraction nor chromatography are required due to the high specificity of the antibody.
After 2 hours incubation at room temperature the microtiter plate is washed to stop the competition reaction.
The substrate solution (tetramethylbenzidine (TMB) – H202) is added and incubated for 30 minutes. The reaction is stopped with H2S04 and the absorbance is measured at the appropriate wavelength. The amount of substrate turnover is inversely proportional to the estradiol concentration in the sample. A calibration curve is plotted and estradiol concentrations in samples are determined by interpolation from the calibration curve.

Storage and Stability: • Store the kit at 2 - 8C until expiration date printed on the kit label.
• Once opened, store the concentrated estradiol-HRP conjugate vial at 2 - 8C. Diluted estradiol-HRP conjugate is stable for 4 hours at room temperature or 24 hours at 2 - 8C when protected from direct exposure to sunlight.
• After reconstitution, store anti-estradiol, calibrators and controls at 2 - 8C for 1 week maximum. For prolonged storage they must be frozen. Three freezing-thawing cycles are allowed.
• Store the unused strips at 2 - 8C in the closed bag containing the desiccant until expiration date.
• The concentrated wash solution is stable at room temperature until expiration date. Prepare fresh diluted wash solution each day.
• The freshly prepared substrate solution is stable for a maximum of 15 minutes at room temperature and must be discarded after use.

Sensitivity and Specificity: The percentage of cross-reaction was estimated under physiological conditions in serum by comparison of the concentration yielding a 50% binding inhibition:

Product Name: Estradiol, 17b (E2)

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