Catalog No: OKAU00028
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OKAU00028
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Datasheets/ManualsPrintable datasheet for Estrone ELISA Kit (OKAU00028)
Product Info
ApplicationEnzyme-linked immunosorbent assay
ELISA Kit LinearityLinearity was determined by taking two urine samples diluted with Assay Buffer, one with a low diluted estrone level of 71.2 pg/mL and one with a higher diluted level of 194.4 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 UrineLow UrineExpected Conc. (pg/mL)Observed Conc. (pg/mL)% Recovery
80%20%169.8157.893.0
60%40%145.1138.395.3
40%60%120.5117.297.3
20%80%95.885.689.3
Mean Recovery93.7%
ELISA Kit PrincipleThe DetectX® Estrone Immunoassay Kit is designed to quantitatively measure estrone present in extracted dried fecal samples, urine and tissue culture media samples. The kit is unique as it measures both non-conjugated estrone and conjugated estrone, in the form of estrone-3-sulfate and estrone 3-glucuronide, in urine and fecal samples with almost equal affinity, allowing for non-invasive testing of this steroid. Please read the complete kit insert before performing this assay. An estrone 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 estrone-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 estrone to each well. After a 2 hour incubation the plate is washed and substrate is added. The substrate reacts with the bound estrone-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 estrone in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.
ELISA Kit ReproducibilityIntra Assay Precision Three urine samples were diluted with Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated Estrone concentrations were:
SampleEstrone Conc. (pg/mL)%CV
1278.14.0
2165.74.4
365.75.7
Inter Assay Precision Three urine samples were diluted with Assay Buffer and run in duplicates in twelve assays run over multiple days by three operators. The mean and precision of the calculated Estrone concentrations were:
SampleEstrone Conc. (pg/mL)%CV
1277.54.2
2168.24.9
364.27.3
ELISA Kit Component
ComponentQuantity
Coated Clear 96 Well Plates1 or 5 Each
Estrone Standard125 uL or 625 uL
DetectX® Estrone Antibody3 mL or 13 mL
DetectX® Estrone Conjugate3 mL or 13 mL
Assay Buffer Concentrate28 mL or 55 mL
Wash Buffer Concentrate30 mL or 125 mL
TMB Substrate11 mL or 55 mL
Stop Solution5 mL or 25 mL
Plate Sealer1 or 5 Each
Additional InformationBackground: Estrone, C18H22O2, also known as E1 or osterone (3-hydroxy-1,3,5(10)-estratrien-17-one) is a C-18 steroid hormone. Estrone is one of the three naturally occurring estrogens, the others being estradiol and estriol. Estrone is produced primarily from androstenedione originating from the gonads or the adrenal cortex and from estradiol by 17-hydroxysteroid dehydrogenase. Androstenedione is also converted into estrone by aromatase (CYP19) to estrone and is expressed in stromal and carcinoma or parenchymal components of breast cancer tissue. Estrone concentrations in premenopausal mammals fluctuate according to the menstrual cycle. In premenopausal women, more than 50% of the estrone is secreted by the ovaries. In prepubertal children, men and non-supplemented postmenopausal women the major portion of estrone is derived from peripheral tissue conversion of androstenedione. Interconversion of estrone and estradiol also occurs in peripheral tissue. In humans, during the follicular phase of the menstrual cycle estrone levels increase slightly. The production of estrone then increases markedly to peak at around day 13. The peak is of short duration and by day 16 the estrone levels will be low. A second peak occurs at around day 21 of the cycle and if fertilization does not occur, then the production of estrone decreases.
::Detection Limit: 28.2 pg/mL
::Cross Reactivity: The following cross reactants were tested in the assay and calculated at the 50% binding point.
SteroidCross Reactivity (%)
Estrone100%
Estrone 3-glucuronide112%
Estrone 3-sulfate65.5%
Estradiol5.0%
Estradiol-3-sulfate<0.1%
Estriol<0.1%
Progesterone<0.1%
Pregnandiol<0.1%
Cortisol< 0.1%
Androsterone< 0.1%
Reconstitution and Storage-20°C
Sample TypeDried Fecal Extracts, Urine and Tissue Culture Media
Sensitivity22.4 pg/mL
Gene Full NameEstrone
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