Estradiol ELISA Kit (OKAU00027)

Catalog No: OKAU00027

ELISA Standard Curve

Product Info

• Application: Enzyme-linked immunosorbent assay
• ELISA Kit Linearity: Linearity was determined by taking two urine samples diluted with Assay Buffer, one with a low diluted estradiol level of 99.6 pg/mL and one with a higher diluted level of 4,098 pg/mL, and mixing them in the ratios given below. The measured concentrations were compared to the expected values based on the ratios used.

  High Urine	Low Urine	Observed Conc. (pg/mL)	Expected Conc. (pg/mL)	% Recovery
  80%	20%	3,393	3,298	102.9
  60%	40%	2,620	2,499	104.9
  40%	60%	1,772	1,699	104.3
  20%	80%	1,021	899	113.6
  			Mean Recovery	106.4%

• ELISA Kit Principle: The DetectX® Estradiol Immunoassay kit uses a specifically generated antibody to measure estradiol and its metabolites in urine and fecal samples. This kit is not recommended for serum, plasma, or saliva samples as the concentration of estradiol in these samples is too low to be measured without significant concentration. The kit will quantitatively measure Estradiol present in reconstituted buffer samples and tissue culture media samples. Please read the complete kit insert before performing this assay. An estradiol standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture rabbit antibodies. An estradiol-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a polyclonal antibody to estradiol to each well. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound estradiol-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. The concentration of the estradiol in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.
• ELISA Kit Reproducibility: Intra Assay Precision Three human samples were diluted with Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated Estradiol concentrations were:

  Sample	Estradiol Conc. (pg/mL)	%CV
  1	2,935	3.9
  2	881.3	4.0
  3	213.9	7.3

  Inter Assay Precision Three human samples were diluted with Assay Buffer and run in duplicates in thirteen assays run over multiple days by four operators. The mean and precision of the calculated Estradiol concentrations were:

  Sample	Estradiol Conc. (pg/mL)	%CV
  1	2,797	3.6
  2	851.2	7.8
  3	216.1	13.8

• ELISA Kit Component:
  • Coated Clear 96 Well Plates: 1 or 5 Each
  • Estradiol Standard: 125 uL or 625 uL
  • DetectX® Estradiol Antibody: 3 mL or 13 mL
  • DetectX® Estradiol Conjugate: 3 mL or 13 mL
  • Assay Buffer Concentrate: 28 mL or 55 mL
  • Wash Buffer Concentrate: 30 mL or 125 mL
  • TMB Substrate: 11 mL or 55 mL
  • Stop Solution: 5 mL or 25 mL
  • Plate Sealer: 1 or 5 Each
• Additional Information: Background: 17b-Estradiol, C18H24O2, also known as E2 or oestradiol (1, 3, 5(10)-Estratrien-3, 17b-diol) is a key regulator of growth, differentiation, and function in a wide array of tissues, including the male and female reproductive tracts, mammary gland, brain, skeletal and cardiovascular systems. The predominant biological effects of E2 are mediated through two distinct intracellular receptors, ERa and ERb, each encoded by unique genes possessing the functional domain characteristics of the steroid/thyroid hormone superfamily of nuclear receptors. ERa is the predominant form expressed in the breast, uterus, cervix, and vagina. ERb exhibits a more limited pattern and is primarily expressed in the ovary, prostate, testis, spleen, lung, hypothalamus, and thymus. Estradiol also influences bone growth, brain development and maturation, and food intake, and it is also critical in maintaining organ functions during severe trauma. In plasma, estradiol is bound to serum proteins such as albumin and sex hormone-binding globulin. Just over 2% of E2 is free and biologically active, the percentage remaining constant throughout the menstrual cycle. Estradiol is conjugated in the liver to sulfate and glucuronide derivatives and excreted. Deactivation includes conversion to less-active estrogens, such as estrone and estriol. Estriol is the major urinary metabolite.
  Detection Limit: 26.5 pg/mL
  Cross Reactivity: The following cross reactants were tested in the assay and calculated at the 50% binding point.

  Steroid	Cross Reactivity (%)
  17b-Estradiol	100%
  Estrone	0.78%
  17a-Estradiol	0.22%
  17a-Ethynylestradiol	0.11%
  Estrone Sulfate	< 0.10%
  Progesterone	< 0.10%
  Testosterone	< 0.10%
  5a-dihydroprogesterone	< 0.10%
  Cortisol	< 0.10%
  Corticosterone	< 0.10%

• Reconstitution and Storage: 2°C to 8°C
• Sample Type: Dried Fecal Extracts, Urine and Tissue Culture Media
• Sensitivity: 39.6 pg/mL

Target Info

• Gene Full Name: Estradiol