|Estrone is a member of estrogen, which is now attracting public attention as an environmental pollutant, especially in rivers and wastewater. Yanaihara Institute Inc. developed a quantitative EIA kit with high specificity and sensitivity for estrone in environmental water. This assay kit is provedto have crossreactivity with neither testosterone nor androstenedione.
More practically, this estrone EIA kit can be used in assessment of endocrine disrupting effects of environmental contaminants and chemicals used in commercial products. In brief, using a human ovarian granulose-like tumor cell line with high aromatase activity and substantially no synthetic activity of androgen and estrogen, the effect of a test compound on the aromatase, a key enzyme in the conversion of androgens to estrogens, is assessed by measuring estrone in culture medium produced from androstenedione added. Reduced concentration of estrone in the medium screens compounds that can disrupt endocrine function by influencing aromatase activity. The usefulness of our estrone specific EIA kit was fully established for this purpose.
This EIA kit is used for quantitative determination of estrone in environmental water and culture medium supernatant. The kit is characterized for sensitive quantification, high specificity and no influence with other components in culture medium supernatant.
The EIA kit shows the following crossreactivities: 100% to estrone, 30.6% to 17β- estradiol and 0.4% to estriol. The crossreactivity to testosterone, progesterone and 4-androstene-3, 17-dione are less than 0.0049%.
This EIA kit for determination of estrone in environmental water and culture supernatant sample is based on a competitive enzyme immunoassay using combination of highly specific antibody to estrone and biotin-avidin affinity system. The 96-wells plate is coated with goat anti rabbit IgG. Biotinylated estrone, estrone standard or samples and rabbit anti estrone antibody are added to the wells for competitive immunoreaction. After incubation and plate washing, HRP labeled streptoavidin (SA-HRP) are added to form HRP labeled streptoavidin-biotinylated estrone-antibody complex on the surface of the wells. Finally, HRP enzyme activity is determined by o-Phenylenediamine dihydrochloride (OPD) and the concentration of estrone is calculated.
|1. Antibody coated plate||MTP*1||1 plate
|Goat anti rabbit IgG|
|2. Standard||lyophilized||1 vial
|3. Labeled antigen||lyophilized||1 vial
|4. Specific antibody||lyophilized||1 bottle||Rabbit anti estrone antibody|
|5. SA-HRP solution||liquid||1 bottle
|HRP labeled streptoavidin|
|6. Substrate buffer||liquid||1 bottle
|0.015% Hydrogen peroxide|
|7. OPD tablet||tablet||2 tablets||o-Phenylenediamine dihydrochloride|
|8. Stopping solution||liquid||1 bottle
|9. Buffer solution||Liquid||1 bottle
|10. Washing solution
|11. Adhesive foil||3 sheets|
MTP*1........ Microtitration plat