Estradiol 17b, Human (17b Estradiol, Estradiol E2, Ethynyl Estradiol, Oestradiol) BioAssay(TM) ELISA Kit

Estradiol (E2) is a C18 steroid hormone with a phenolic A ring. This steroid hormone has a molecular weight of 272.4. It is the most potent natural Estrogen, produced mainly by the ovary, placenta, and in smaller amounts by the adrenal cortex, and the male testes (1,2,3). Estradiol (E2) is secreted into the blood stream where 98% of it circulates bound to sex hormone binding globulin (SHBG). To a lesser extent it is bound to other serum proteins such as albumin. Only a tiny fraction circulates as free hormone or in the conjugated form (4,5). Estrogenic activity is effected via estradiol-receptor complexes which trigger the appropriate response at the nuclear level in the target sites. These sites include 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 (6,7). The rising estradiol concentration is understood to exert a positive feedback influence at the level of the pituitary where it influences the secretion of the gonadotropins, follicle stimulating hormone (FSH), and luteinizing hormone (LH), which are essential for follicular maturation and ovulation, respectively (8,9). Following ovulation, estradiol levels fall rapidly until the luteal cells become active resulting in a secondary gentle rise and plateau of estradiol in the luteal phase. During pregnancy, maternal serum Estradiol levels increase considerably, to well above the pre-ovulatory peak levels and high levels are sustained throughout pregnancy (10). Serum Estradiol measurements are a valuable index in evaluating a variety of menstrual dysfunctions such as precocious or delayed puberty in girls (11) and primary and secondary amenorrhea and menopause (12). Estradiol levels have been reported to be increased in patients with feminizing syndromes (14), gynaecomastia (15) and testicular tumors (16). In cases of infertility, serum Estradiol measurements are useful for monitoring induction of ovulation following treatment with, for example, clomiphene citrate, LH-releasing hormone (LH-RH), or exogenous gonadotropins (17, 18). During ovarian hyperstimulation for in vitro fertilization (IVF), serum estradiol concentrations are usually monitored daily for optimal timing of human chorionic gonadotropin (hCG) administration and oocyte collection (19). \n\nSample Type:\nSerum\n\nIntended Use: \nFor the quantitative determination of Estradiol (E2) concentration in human serum.\n\nTest Principle: \nThe E2 EIA is based on the principle of competitive binding between E2 in the test specimen and E2-HRP conjugate for a constant amount of rabbit anti-Estradiol. In the incubation, goat anti-rabbit IgG-coated wells are incubated with 25ul E2 standards, controls,\npatient samples, 100ul Estradiol-HRP Conjugate Reagent and 50ul rabbit anti-Estradiol reagent at room temperature (18-25 degrees C) for 90 minutes. 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. Thus, the amount of E2 peroxidase conjugate immunologically bound to the well progressively decreases as the concentration of E2 in the specimen increases. Unbound E2 peroxidase conjugate is then removed and the wells 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 1N HCl, and the absorbance is measured spectrophotometrically at 450nm. The intensity of the color formed is proportional to the amount of enzyme present and is inversely related to the amount of unlabeled E2 in the sample. A standard curve is obtained by plotting the concentration of the standard versus the absorbance. The E2 concentration of the specimens and controls run concurrently with the standards can be calculated from the standard curve.\n\nKit Components:\nMicrotiter, 1x96 wells, Goat Anti-Rabbit IgG-coated \nStandard, 0pg/ml, 1x500ul\nStandard, 10pg/ml, 1sx500ul\nStandard, 30pg/ml, 1x500ul\nStandard, 100pg/ml, 1x500ul\nStandard, 300pg/ml, 1x500ul\nStandard, 1000pg/ml, 1x500ul \nRabbit Anti-Estradiol Reagent (pink color), 1x7ml\nEstradiol-HRP Conjugate Reagent (blue color), 1x12ml\nEstradiol Control 1, Liquid, 0.5 ml, Ready to use.\nEstradiol Control 2, Liquid, 0.5 ml, Ready to use.\nTMB Reagent (One-Step), 1x11ml.\nStop Solution (1N HCl), 1x11ml.\n\nStorage and Stability:\nStore all components at 4 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

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SPECIFICATIONS

Catalog Number

E3550-09C

Size

1Kit

References

1. Tsang, B.K., Armstrong, D.T. and Whitfield, J.F., Steroid biosynthesis by isolated human ovarian follicular cells in vitro, J. Clin. Endocrinol. Metab., 1980; 51: 1407-1411. 2. Gore-Langton, R.E. and Armstrong, D.T., Follicular steroidogenesis and its control. In: Knobil, E., and Neill, J. et al., ed. The Physiology of Reproduction. Raven Press, New York; 1988: 331-385. 3. Hall, P.F., Testicular steroid synthesis: organization and regulation. In: Knobil, E., and Neill, J. et al., ed. The Physiology of Reproduction. Raven Press, New York; 1988: 975-998. 4. Siiteri, P.K., Murai, J.T., Hammond, G.L., Nisker, J.A., Raymoure, W.J. and Kuhn, R.W., The serum transport of steroid hormones, Rec. Prog. Horm. Res., 1982; 38: 457-510.\n5. Baird, D.T., Ovarian steroid secretion and metabolism in women. In: James, V.H.T., Serio, M. and Giusti, G., eds. The Endocrine Function of the Human Ovary. Academic Press, New York; 1976: 125-133.\n6. McNatty, K.P., Baird, D.T., Bolton, A., Chambers, P., Corker, C.S. and McLean, H., Concentration of oestrogens and androgens in human ovarian venous plasma and follicular fluid throughout the menstrual cycle, J. Endocrinol., 1976; 71: 77- 85. 7. Abraham, G.E., Odell, W.D., Swerdloff, R.S., and Hopper, K.,\nSimultaneous radioimmunoassay of plasma FSH, LH, progesterone, 17-hydroxyprogesterone and estradiol-17b during the menstrual cycle, J. Clin. Endocrinol. Metab., 1972; 34: 312-318. 8. March, C.M., Goebelsmann, U., Nakumara, R.M., and Mishell, D.R. Jr., Roles of estradiol and progesterone in eliciting the\nmidcycle luteinizing hormone and follicle-stimulating hormone surges. J. Clin. Endocrinol. Metab., 1979; 49: 507-513. 9. Simpson, E.R., and MacDonald, P.C., Endocrinology of pregnancy. In: Williams, R.H., ed., Textbook of Endocrinology. Saunders Company, Philadelphia; 1981: 412-422. 10. Jenner, M.R., Kelch, R.P., Kaplan, S.L. and Grumbach, M.M., Hormonal changes in puberty: IV. Plasma estradiol, LH, and FSH in prepubertal children, pubertal females and in precocious puberty, premature thelarche, hypogonadism and\nin a child with feminizing ovarian tumor. J. Clin. Endocrinol. Metab., 197 2; 34: 521-530. 11. Goldstein, D., Zuckerman, H., Harpaz, S., et al., Correlation between estradiol and progesterone in cycles with luteal phase deficiency. Fertil. Steril., 1982; 37: 348-354. 12. Kirschner, M.A., The role of hormones in the etiology of human breast cancer. Cancer, 1977; 39: 2716-2726. 13. Odell, W.D. and Swerdloff, R.S., Abnormalities of gonadal function in men. Clin. Endocr., 1978; 8: 149-180. 14. MacDonald, P.C., Madden, J.D., Brenner, P.F., Wilson, J.D. and Siiteri, P.K., Origin of estrogen in normal men and in women with testicular feminization, J.Clin. Endocrinol. Metabl., 1979; 49: 905-916. 15. Fishel, S.B., Edwards, R.G., Purdy, J.M., Steptoe, P.C., Webster, J., Walters, E., Cohen, J., Fehilly, C. Hewitt, J., and Rowland, G., Implantation, abortion and birth after in vitro fertilization using the natural menstrual cycle or follicular stimulation with clomiphene citrate and human menopausal gonadotropin, J. In Vitro Fertil. Embryo Transfer, 1985; 2: 123- 131. 16. Ratcliffe, W.A., Carter, G.D., Dowsett, M., et al., Oestradiol assays: applications and guidelines for the provision of a clinical biochemistry service, Ann. Clin. Biochem., 1988;\n25:466-483. 17. Tietz, N.W. ed., Clinical Guide to Laboratory Tests, 3rd Edition, W.B. Saunders, Co., Philadelphia, 1995: 216-217. 18. USA Center for Disease Control/National Institute of Health\nManual,