Calbiotech Estradiol ELISA

SUMMARY AND EXPLANATION OFTHE TEST 
Estradiol E2 is the most potent natural Estrogen, produced mainly by the ovary, placenta, and in smaller amounts by the adrenal cortex, and the male testes. Estradiol is secreted into the blood stream where 98% bound to sex hormone binding globulin (SHBG). Estrogenic activity is effected via estradiol-receptor complexes which trigger the appropriate response at the follicles, uterus, breast, vagina, urethra, hypothalamus, pituitary and to a lesser extent the liver and skin. In non-pregnant women with normal menstrual cycles, estradiol secretion follows a cyclic, biphasic pattern with the highest concentration found immediately prior to ovulation. During pregnancy, maternal serum Estradiol levels increase considerably, to well above the pre-ovulatory peak levels and high levels are sustained throughout pregnancy. Serum Estradiol measurements are a valuable index in evaluating a variety of menstrual dysfunctions such as precocious or delayed puberty in girls and primary and secondary amenorrhea and menopause. Estradiol levels have been reported to be increased in patients with feminizing syndromes, gynaecomastia and testicular tumors. In cases of infertility, serum Estradiol measurements are useful for monitoring induction of ovulation following treatment.

PRINCIPLE OF THE TEST 
The Calbiotech, Inc E2 ELISA kit is based on the principle of Delayed competitive binding assay between E2 in the test specimen and E2 enzyme conjugate for a constant amount of anti-Estradiol monoclonal antibody epitops (Biotin reagent). In the incubation, anti-E2 antibody biotin reagent, E2 standards, controls, and samples are incubated for 45 minutes at room temperature (RT), then E2 enzyme conjugate is added on the top of the reaction mixture and incubation continues for 45 minutes more. During the incubation, a fixed amount of HRP-labeled E2 competes with the endogenous E2 in the standard, sample, or quality control serum for a fixed number of binding sites of the specific E2 antibody. E2 peroxidase conjugate immunologically bound to the well progressively decreases as the concentration of E2 in the specimen increases. Unbound of anti-Estradiol Biotin Reagent and E2 peroxidase conjugate is then removed and the wells are washed. Next, a solution of TMB Reagent is added and incubated at room temperature for 20 minutes, resulting in the development of blue color. The color development is stopped with the addition of stop solution, and the absorbance is measured spectrophotometrically at 450 nm. A standard curve is obtained by plotting the concentration of the standard versus the absorbance.

STORAGE AND STABILITY 
1. Store the kit at 2-8°C. 
2. Keep microwells sealed in a dry bag with desiccants. 
3. The reagents are stable until expiration of the kit. 
4. Do not expose test reagents to heat, sun or strong light