The United States Biological Human Estradiol-17b (E2) ELISA kit is a competitive binding immunoassay for the quantitative determination of estradiol in serum and plasma.
A fixed amount of estradiol labeled with horseradish peroxidase (HRP) competes with unlabeled estradiol present in standards or samples for a limited number of binding sites of a specific antibody. The E2-HRP-antibody complex is simultaneously fixed on the wells of the microtiter plate coated with an excess of anti-rabbit gamma-globulins. Neither extraction nor chromatography are required due to the high specificity of the antibody. After 2 hours incubation at rooom temperature the microtiter plate is washed to stop the competition reaction. The substrate solution (TMB-H2O2) is added and incubated for 30 minutes. The reaction is stopped with sulfuric acid solution and the absorbance is measured at the appropriate wavelength. The amount of substrate turnover is inversely proportional to the estradiol concentration in the sample. A standard curve is plotted and estradiol concentrations in samples are determined by interpolation from the standard curve.
Sample Type:Serum or plasma, 50ul sample volume required.
Sensitivity:5 ± 2pg/ml (mean ± SD)
Range: 10-1000pg/ml
Incubation Time:2 hours 30 minutes at room temperature.
Kit Components:E3550-06A: Standard 1, 0pg/ml, 1x1vialE3550-06B: Standard 2, 30pg/ml, 1x1vialE3550-06C: Standard 3, 56pg/ml, 1x1vialE3550-06D: Standard 4, 136pg/ml, 1x1vialE3550-06E: Standard 5, 283pg/ml, 1x1vialE3550-06F: Standard 6, 995pg/ml, 1x1vialE3550-06G: Microtiter Plate, 1x96 wellsE3550-06H: Estradiol Ab, 1x1 vialE3550-06J: Low Control, 93+22pg/mlE3550-06K: High Control, 206+40pg/mlE3550-06L: Estradiol Ab (HRP), 1x500ulE3550-06M: Conjugate Buffer, 3x6mlE3550-06N: Wash Buffer, 1x10mlE3550-06P: TMB, 1x1mlE3550-06Q: Substrate Buffer, 3x21mlE3550-06R: Stop Solution, 1x6ml
Storage and Stability:Store other components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
