GenWay/Estradiol (E2) Enzyme Immunoassay Test Kit/GWB-AFC61F/ELISA_Kits

Description:

Species: Human.

Sample Type: Serum.

For the quantitative determination of Cardiac-Specific Troponin-I in serum. Blood, Genitalia, Endocrine Glands, Cells Cultured, Neoplasms by Site Please click here for MSDS PDF Datasheet.

Product Name: Estradiol (E2) Concentration.

Sample Type: Serum.

# of Samples: 1X96 Assays.

Intended Use: For the quantitative determination of Estradiol (E2) concentration in human serum.

Introduction: 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).

Principle of the test: The 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 25 ?l E2 standards, controls, patient samples, 100 ?l Estradiol-HRP Conjugate Reagent and 50 ?l rabbit anti-Estradiol reagent at room temperature (18-25C) 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 450 nm. 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.

Limitation of the Test:
1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the package insert instructions and with adherence to good laboratory practice.
2. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
3. Serum samples demonstrating gross lipemia, gross hemolysis, or turbidity should not be used with this test.
4. The results obtained from the use of this kit should be used only as an adjunct to other diagnostic procedures and information available to the physician.

Product Name: Estradiol (E2) Concentration.

Function: The branched-chain alpha-keto dehydrogenase complex catalyzes the overall conversion of alpha-keto acids to acyl-CoA and CO(2). It contains multiple copies of three enzymatic components: branched-chain alpha-keto acid decarboxylase (E1), lipoamide acyltransferase (E2) and lipoamide dehydrogenase (E3).

Catalytic Activity: 2-methylpropanoyl-CoA + enzyme N6-(dihydrolipoyl)lysine = CoA + enzyme N6-(S-(2-methylpropanoyl)dihydrolipoyl)lysine.

Cofactor: Binds 1 lipoyl cofactor covalently.

Subunit: Forms a 24-polypeptide structural core with octahedral symmetry.

Subcellular Location: Mitochondrion matrix.

Disease: Defects in DBT are the cause of maple syrup urine disease type 2 (MSUD2) [MIM:248600]. MSUD is an autosomal recessive disorder characterized by mental and physical retardation, feeding problems, and a maple syrup odor to the urine.

Miscellaneous: The catalytic function of this enzyme is to accept, and to transfer to coenzyme A, acyl groups that are generated by the branched-chain alpha-keto acid decarboxylase component.

Similarity: Belongs to the 2-oxoacid dehydrogenase family.

Similarity: Contains 1 lipoyl-binding domain.

Product Protocol: To see this product used in a peer reviewed journal, click the following link:
Proteomic Signatures in Plasma during Early Acute Renal Allograft Rejection

Additional Information: