CLIA Kit for Estradiol (E2)

17B-Estradiol; Oestradiol; Beta-Estradiol

Specificity

This assay has high sensitivity and excellent specificity for detection of Estradiol (E2).
No significant cross-reactivity or interference between Estradiol (E2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Estradiol (E2) and the recovery rates were calculated by comparing the measured value to the expected amount of Estradiol (E2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 85-101 92
EDTA plasma(n=5) 87-104 93
heparin plasma(n=5) 90-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Estradiol (E2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Estradiol (E2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Estradiol (E2) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 89-96% 91-105% 98-105% 82-98%
EDTA plasma(n=5) 82-104% 92-101% 83-97% 80-103%
heparin plasma(n=5) 85-92% 88-99% 79-105% 90-98%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Substrate A 1×10mL Substrate B 1×2mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 100µL Substrate Solution. Incubate 10 minutes at 37°C;
7. Read RLU value immediately.

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Magazine Citations
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Fertility and Sterility Growth differential factor-9 inhibits testosterone production in mouse theca interstitial cells Pubmed: 23954356
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Academic Sciences Piper longum hexane fraction induces infertility by modulation of inflammatory mediators and gonadotropin insufficiency in female rats Ijppsjournal: Source
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Reproductive Toxicology Carbamazepine-exposure during gestation and lactation affects pubertal onset and spermatic parameters in male pubertal offspring Pubmed:24126190
Phytomedicine. Phloretin promotes osteoclast apoptosis in murine macrophages and inhibits estrogen deficiency-induced osteoporosis in mice Pubmed:24932975
Turkish Journal of Urology Apoptosis in testicular tissue of rats after vasectomy: evaluation of eNOS, iNOS immunoreactivities and the effects of ozone therapy Turkishjournalofurology:Source
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Reprod Toxicol In utero PubMed: 25530038
oncology research Mechanistic Study on Triptorelin Action in Protecting From 5-FU-Induced Ovarian Damage in Rats PubMed: 26629940
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Archives of Physiology and Biochemistry Activation of cardiac renin–angiotensin system and plasminogen activator inhibitor-1 gene expressions in oral contraceptive-induced cardiometabolic disorder Pubmed:26934364
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Medical Science Monitor Up-Regulation of Long Noncoding RNA SRA Promotes Cell Growth, Inhibits Cell Apoptosis, and Induces Secretion of Estradiol and Progesterone in Ovarian … Pubmed:29674607
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