Measures Estradiol levels in various samples. This assay cross-reacts 0.07% with 17 alpha -ethynylestradiol, 0.03% with 17 beta -estradiol 3-Benzoate, 0.86% with Estriol, 0.26% with Estrone and 0.06% with Progesterone.
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference observed with 1 or more available related molecules.
The Parameter Estradiol Immunoassay is a 3.5 hour competitive enzyme
immunoassay designed to measure Estradiol in cell culture supernates, serum, and plasma.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in twenty separate
assays to assess inter-assay precision. Assays were performed by at
least three technicians using two lots of kit components.
The recovery of Estradiol spiked to levels throughout the range of
the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high
concentrations of Estradiol were serially diluted with the appropriate
Calibrator Diluent to produce samples with values within the dynamic
range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Estradiol Primary Antibody Solution
Add 100 µL of Estradiol Primary Antibody Solution to each well (excluding the NSB wells). Cover with a plate sealer, and incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, control, or sample to the appropriate wells.
100 µL Calibrator Diluent RD5-62
Add 100 µL of Calibrator Diluent RD5-62 to the zero standard (B0) wells and NSB wells.
50 µL Estradiol Conjugate
Add 50 µL of Estradiol Conjugate to all wells. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
100 µL Stop Solution
Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.