17 Porcine -Estradiol

Brand

GENWAY

Overview

Intended Use: Competitive immunoenzymatic colorimetric method for quantitative determination of 17 β -Estradiol in human serum or plasma. Introduction: Estradiol(17β-Estradiol) is a sex hormone. It represents the major Estrogen in humans. Estradiol has not only a critical impact on reproductive and sexual functioning but also affects other organs including bone structure. During the reproductive years most Estradiol in women is produced by the ovaries smaller amounts of Estradiol are also produced by the adrenal cortex. In men the testes produce Estradiol. In plasma Estradiol is largely bound to sex hormone binding globulin (SHBG) also to albumin only a fraction is free and biologically active. Serum Estradiol measurement in women reflects primarily the activity of the ovaries. During pregnancy Estrogen levels including Estradiol rise steadily towards term. Estradiol increases due to placental production. In adult premenopausal women ovarian production of Estradiol is stimulated by luteinizing hormone (LH) and the follicle-stimulating hormone (FSH) during the menstrual cycle. In adult women Estradiol levels are measured in the evaluation of fertility and menstrual irregularities and to monitor ovarian follicular function during induction of ovulation. In the female Estradiol acts as a growth hormone for tissue of the reproductive organs. The development of secondary sexual characteristics in women is driven by Estradiol. Estradiol is involved also in men fertility. Estradiol regulates the bone maintenance. Post-menopause women experience an accelerated los of bone mass due to a relative Estrogen deficiency. Estradiol affects the production of multiple proteins including lipoproteins binding proteins and proteins responsible for blood clotting. Estrogens have been found to have neuroprotective function. The Estradiol for his activities is involved in some types of cancer such as breast cancer and cancer of the uterine lining. In addition there are several benign gynaecologic conditions that are dependent on Estrogen such as endometriosis leiomyomata uteri and uterine bleeding. Principles of the assay: Microtiter strip wells are precoated with anti-Estradiol antibodies (solid-phase). Estradiol in the sample competes with added horseradish peroxidase labelled Estradiol (enzyme-labelled antigen) for antibody binding. After incubation a bound/free separation is performed by solid-phase washing. The immune complex formed by enzyme-labelled antigen is visualized by adding Tetramethylbenzidine (TMB) substrate which gives a blue reaction product. The intensity of this product is inversely proportional to the amount of Estradiol in the sample. Sulphuric acid is added to stop the reaction. This produces a yellow endpoint colour. Absorption at 450 nm is read using an ELISA microwell plate reader. Storage and Stability: The reagents are stable up to the expiry date stated on the label when stored at 2. . . 8 °C in the dark. Limitations of the Test: Sample(s) which are contaminated microbiologically should not be used in the assay. Highly lipemic or haemolysed specimen(s) should similarly not be used. It is important that the time of reaction in each well is held constant for reproducible results. Pipetting of samples should not extend beyond ten minutes to avoid assay drift. If more than one plate is used it is recommended to repeat the dose response curve. Addition of the substrate solution initiates a kinetic reaction which is terminated by the addition of the stop solution. Therefore the addition of the substrate and the stopping solution should be added in the same sequence to eliminate any time deviation during reaction. Plate readers measure vertically. Do not touch the bottom of the wells. Failure to remove adhering solution adequately in the aspiration or decantation wash step(s) may result in poor replication and spurious results. References Joshi U. M. et al. (1979) Steroids 34(1) 35Exley D. and Abuknesha R. (1978) Febs Letters 91(2) 162Ismail A. A. Niswender G. D. and Midgley A. R. (1972) J. Clin:Endocrin. Metab 34 177-184Rajkowski K. M. Cittanova N. Desfosses B. and Jayle M. F. (1977) Steroids 29 (5) 701-713Sadem D. Sela E. and Hexter C. S. (1979) J. of Immunological Methods 28 125-131Wisdom G. B. (1976) Clin Chem. 22 (8) 1243-1255

spezie

other

detection target

17 Porcine -Estradiol

pack size

1x96 assay