Estradiol Parameter Assay Kit

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Multi-species Estradiol ELISA Standard Curve
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Product Details
Citations (14)
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Estradiol Parameter Assay Kit Summary

96-well strip plate
Assay Length
3.5 hours
12.1 pg/mL
Assay Range
12.3 - 3,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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.

Product Summary

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

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 153 744 1419 164 765 1533
Standard Deviation 13.6 26.8 76.6 9.54 61.5 116
CV% 8.9 3.6 5.4 5.8 8 7.6


The recovery of Estradiol spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 94 81-102
EDTA Plasma (n=4) 103 90-113
Heparin Plasma (n=4) 102 88-117
Serum (n=4) 96 87-108


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.
Multi-species Estradiol ELISA Linearity

Data Examples

Multi-species Estradiol ELISA Standard Curve

Product Datasheets

Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to

Assay Procedure

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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Estradiol Primary Antibody Solution
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  6. 100 µL Standard, Control, or Sample
  7.   Add 100 µL of Standard, control, or sample to the appropriate wells.

  8. 100 µL Calibrator Diluent RD5-62
  9.   Add 100 µL of Calibrator Diluent RD5-62 to the zero standard (B0) wells and NSB wells.

  10. 50 µL Estradiol Conjugate
  11.   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.
  12.   Aspirate and wash 4 times.

  13. 200 µL Substrate Solution
  14.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  15. 100 µL Stop Solution
  16.   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.

Citations for Estradiol Parameter Assay Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

14 Citations: Showing 1 - 10
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  1. Beneficial effects of a decreased meal frequency on nutrient utilization, secretion of luteinizing hormones and ovarian follicular development in gilts
    Authors: L Hua, L Zhao, Z Mao, W Li, J Li, X Jiang, L Che, S Xu, Y Lin, Z Fang, B Feng, Wu, Y Zhuo
    Journal of animal science and biotechnology, 2021;12(1):41.
    Species: Bovine
    Sample Types: Serum
  2. A prolonged hiatus in postmenopausal HRT, does not nullify the therapy's positive impact on ageing related sarcopenia
    Authors: GL Onambélé-P, DJ Tomlinson, CI Morse, H Degens
    PLoS ONE, 2021;16(5):e0250813.
    Species: Human
    Sample Types: Serum
  3. Adenine-induced chronic kidney disease induces a similar skeletal phenotype in male and female C57BL/6 mice with more severe deficits in cortical bone properties of male mice
    Authors: CE Metzger, EA Swallow, AJ Stacy, MR Allen
    PLoS ONE, 2021;16(4):e0250438.
    Species: Mouse
    Sample Types: Serum
  4. Circulating Endothelial Progenitor Cells Are Preserved in Female Mice Exposed to Ambient Fine Particulate Matter Independent of Estrogen
    Authors: X Liu, Y Xiao, Q Zhu, Y Cui, H Hao, M Wang, PJ Cowan, RJ Korthuis, G Li, Q Sun, Z Liu
    International Journal of Molecular Sciences, 2021;22(13):.
    Species: Mouse
    Sample Types: Serum
  5. Loss of Estrogen Receptors is Associated with Increased Tumor Aggression in Laryngeal Squamous Cell Carcinoma
    Authors: A Verma, N Schwartz, DJ Cohen, V Patel, B Nageris, G Bachar, BD Boyan, Z Schwartz
    Sci Rep, 2020;10(1):4227.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Live Birth in Sex-Reversed XY Mice Lacking the Nuclear Receptor Dax1
    Authors: I Fernandes-, A Milona, KG Murphy, WS Dhillo, BM Owen
    Sci Rep, 2020;10(1):1703.
    Species: Mouse
    Sample Types: Plasma
  7. CB1 agonism prolongs therapeutic window for hormone replacement in ovariectomized mice
    Authors: K Zhang, Q Yang, L Yang, YJ Li, XS Wang, YJ Li, RL Dang, SY Guan, YY Guo, T Sun, YM Wu, A Liu, Y Zhang, SB Liu, MG Zhao
    J. Clin. Invest., 2019;130(0):.
    Species: Human
    Sample Types: Plasma
  8. Experimental study on mandibular length and facial symmetry of low estrogen level and anterior disc displacement of temporomandibular joint
    Authors: Z Jiao, X Wang, X Zhang, C Yang
    Sci Rep, 2018;8(1):15635.
    Species: Rabbit
    Sample Types: Serum
  9. Impairment of learning and memory of mice offspring at puberty, young adulthood, and adulthood by low-dose Cd exposure during pregnancy and lactation via GABAAR ?5 and ? subunits
    Authors: Q Zhao, L Gao, Q Liu, Y Cao, Y He, A Hu, W Chen, J Cao, C Hu, L Li, F Tao
    Ecotoxicol. Environ. Saf., 2018;166(0):336-344.
    Species: Mouse
    Sample Types: Serum
  10. Response of Sclerostin and Bone Turnover Markers to High Intensity Interval Exercise in Young Women: Does Impact Matter?
    Authors: R Kouvelioti, N Kurgan, B Falk, WE Ward, AR Josse, P Klentrou
    Biomed Res Int, 2018;2018(0):4864952.
    Species: Human
    Sample Types: Serum
  11. Effects of Microcystis on Hypothalamic-Pituitary-Gonadal-Liver Axis in Nile Tilapia (Oreochromis niloticus)
    Authors: J Chen, S Meng, H Xu, Z Zhang, X Wu
    Bull Environ Contam Toxicol, 2017;0(0):.
    Species: Fish - Oreochrmois niloticus (Tilapia)
    Sample Types: Serum
  12. Reproductive Hormones and Their Receptors May Affect Lung Cancer
    Authors: M Dou, K Zhu, Z Fan, Y Zhang, X Chen, X Zhou, X Ding, L Li, Z Gu, M Guo, M Yan, X Deng, P Shen, S Wang
    Cell. Physiol. Biochem., 2017;44(4):1425-1434.
    Species: Mouse
    Sample Types: Serum
  13. Phenylethanoid Glycosides of Cistanche on menopausal syndrome model in mice
    Authors: S Tian, M Miao, M Bai, Z Wei
    Saudi Pharm J, 2017;25(4):537-547.
    Species: Mouse
    Sample Types: Serum
  14. Evaluation of Effects of Topical Estradiol Benzoate Application on Cutaneous Wound Healing in Ovariectomized Female Mice
    PLoS ONE, 2016;11(9):e0163560.
    Species: Mouse
    Sample Types: Plasma


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Estradiol Parameter Assay Kit
By Anonymous on 05/02/2018
Application: Sample Tested: FaDu human squamous cell carcinoma cell line