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Estradiol Parameter Assay Kit

R&D Systems | Catalog # KGE014

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Key Product Details

Assay Length

3.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL)

Sensitivity

12.1 pg/mL

Assay Range

12.3-3000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
View all Estradiol ELISA Kits »

Product Summary for Estradiol Parameter Assay Kit

The Parameter Estradiol Immunoassay is a 3.5 hour competitive enzyme immunoassay designed to measure Estradiol in cell culture supernates, serum, and plasma.

Product Specifications

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

Biotin

Species

Multi-Species

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.

精度

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, EDTA Plasma, Heparin Plasma, Serum

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 8.0 7.6

Recovery for Estradiol Parameter Assay Kit

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.

Multi-species Estradiol ELISA Linearity

Scientific Data Images for Estradiol Parameter Assay Kit

Multi-species Estradiol ELISA Standard Curve

Multi-species Estradiol ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Estradiol

Alternate Names

Estrogen

Additional Estradiol Products

Product Documents for Estradiol Parameter Assay Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Estradiol Parameter Assay Kit

For research use only

⚠ 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 www.P65Warnings.ca.gov.

Citations for Estradiol Parameter Assay Kit

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    Sample Tested: FaDu human squamous cell carcinoma cell line
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Protocols

View specific protocols for Estradiol Parameter Assay Kit (KGE014):

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
  3.   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.
  4.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    5. 100 µL Standard, Control, or Sample
  5.   Add 100 µL of Standard, control, or sample to the appropriate wells.
    6. 100 µL Calibrator Diluent RD5-62
  6.   Add 100 µL of Calibrator Diluent RD5-62 to the zero standard (B0) wells and NSB wells.
    7. 50 µL Estradiol Conjugate
  7.   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.
  8.   Aspirate and wash 4 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    10. 100 µL Stop Solution
  10.   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.

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