Estradiol Parameter Assay Kit

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Multi-species Estradiol ELISA Standard Curve
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Product Details
Citations (20)
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Estradiol Parameter Assay Kit Summary

96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
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

Scientific Data

Multi-species Estradiol ELISA Standard Curve

Product Datasheets

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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.

20 Citations: Showing 1 - 10
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  1. Upregulation of the oestrogen target gene SIX1 is associated with higher growth speed and decreased survival in HCV-positive women with hepatocellular carcinoma
    Authors: RM Critelli, F Milosa, A Romanzi, S Lasagni, G Marcelli, L Di Marco, A Pivetti, F Schepis, D Romagnoli, S Mancarella, F Dituri, ML Martinez-C, G Giannelli, E Villa
    Oncology Letters, 2022;24(5):395.
    Species: Human
    Sample Types: Serum
  2. Impact of age and sex on hyperoxia-induced cardiovascular pathophysiology
    Authors: R Vichare, F Saleem, H Mansour, K Bojkovic, F Cheng, M Biswal, SK Panguluri
    Mechanisms of Ageing and Development, 2022;208(0):111727.
    Species: Human
    Sample Types: Serum
  3. Mesh-Tissue Integration of Platelet-Rich Plasma-Decellularized Amnion Scaffold-Polypropylene Mesh Sandwiches Implanted in the Vesicovaginal Spaces of Hypoestrogenic Rabbit Models: Protocol for a Randomized Controlled Trial
    Authors: AP Meutia, BI Santoso, A Hestiantor, PE Wuyung, J Prihartono, A Boediono, S Djusad, A Fauzi, P Budinurdja
    JMIR research protocols, 2022;11(8):e37942.
    Species: Rabbit
    Sample Types: Serum
  4. Comparison of the Toxicological Effects of Pesticides in Non-Tumorigenic MCF-12A and Tumorigenic MCF-7 Human Breast Cells
    Authors: L Coppola, S Tait, E Fabbrizi, M Perugini, C La Rocca
    International Journal of Environmental Research and Public Health, 2022;19(8):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Androgens increase excitatory neurogenic potential in human brain organoids
    Authors: I Kelava, I Chiaradia, L Pellegrini, AT Kalinka, MA Lancaster
    Nature, 2022;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. 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
  7. Identification of the Active Ingredient and Beneficial Effects of Vitex rotundifolia Fruits on Menopausal Symptoms in Ovariectomized Rats
    Authors: JH Lee, S Lee, QN Nguyen, HM Phung, MS Shin, JY Kim, H Choi, SH Shim, KS Kang
    Biomolecules, 2021;11(7):.
    Species: Rat
    Sample Types: Plasma
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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
  18. 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
  19. 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
  20. 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
Sample Tested: FaDu human squamous cell carcinoma cell line