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ELISA Kit for Estrone (E1)
Synonyms: Oestrone
Application:Enzyme-linked immunosorbent assay for Antigen Detection
Research area:Metabolic pathway
Test method:Competitive Inhibition
Shelf life:Reproductive science
Storage conditions:Genetic science
Delivery conditions:Hormone metabolism
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Estrone (E1) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Estrone (E1) and unlabeled Estrone (E1) (Standards or samples) with the pre-coated antibody specific to Estrone (E1). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Estrone (E1) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Estrone (E1) in the sample.
Sensitivity:The minimum detectable dose of this kit is typically less than 8.86pg/mL
Sample type:
plasma and other biological fluids
Assay length:2h
This assay has high sensitivity and excellent specificity for detection of Estrone (E1).
No significant cross-reactivity or interference between Estrone (E1) and analogues was observed.
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Estrone (E1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Estrone (E1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
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.
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 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
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