The Calbiotech Mouse/Rat Estradiol (E2) ELISA Kit is intended for the quantitative determination of Estradiol (E2) concentration in Mouse/Rat serum and plasma.
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 Mouse/Rat E2 ELISA kit is based on the principle of competitive binding between E2 in the test specimen and E2 enzyme conjugate for a constant amount of anti-Estradiol polyclonal antibody. In the incubation, anti-E2 antibody coated wells are incubated with E2 standards, controls, samples, and E2 enzyme conjugate at room temperature for 120 minutes with. 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 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 15 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.
Publications Citing the Calbiotech Mouse/Rat Estradiol ELISA
In summary, the Calbiotech Kit was the only available assay tested that demonstrated good E2 parallelism to the assay standard curve and accuracy vs a gold standard method (i.e. GC/MSMS). Also of note, the Calbiotech assay is sensitive and requires minimal sample volume. Therefore, based on these findings the Calbiotech E2 assay has been implemented for use in mouse serum samples within the Ligand Core.Daniel J. Haisenleder et. al. (See #1 below)