Estradiol Parameter Assay Kit

(2 citations)   
  • Format
    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)
  • Sensitivity
    12.1 pg/mL
  • Assay Range
    12.3 - 3,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    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.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
QC87, Parameter Immunoassay Control Set 791 for Estradiol - Please Inquire
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.

Precision
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
Sample123123
n202020202020
Mean15374414191647651533
Standard Deviation13.626.876.69.5461.5116
CV%8.93.65.45.887.6

Recovery

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
Linearity
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.
Estradiol Parameter Assay Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
  • Alternate Names
    Estradiol; Estrogen;
Related Research Areas
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:

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.

2 Citations: Showing 1 - 2
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Species
Sample Type
  1. 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 Type: Serum
  2. 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 Type: Plasma
ELISA Controls
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Parameter Immunoassay Control Set 791 for Estradiol

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