Human Cortisol ELISA Kit
SKU: E-OSEL-H0006
Human Cortisol ELISA Kit
| Detection Range | 6.25-400 ng/mL |
| Sensitivity | 2.87 ng/mL |
Product Details
Properties
| Assay type | Competitive-ELISA |
| Format | 96T |
| Assay time | 1.5h |
| Detection range | 6.25-400 ng/mL |
| Sensitivity | 2.87 ng/mL |
| Sample type &Sample volume | serum, plasma, urine, saliva; 50μL |
| Specificity | This kit recognizes Cortisol in samples. No significant cross-reactivity or interference between Cortisol and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are < 10%. |
| Application | This ELISA kit applies to the in vitro quantitative determination of Cortisol concentrations in serum, plasma, urine, saliva. |
Test Principle
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Human Cortisol. Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for Human Cortisol are added to the micro ELISA plate wells. Human Cortisol in samples (or standards) competes with a fixed amount of Cortisol on the solid phase supporter for sites on the HRP linked detection antibody specific to Cortisol. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The concentration of Human Cortisol in the samples is then determined by comparing the OD of the samples to the standard curve.
Kit Components and Storage
An unopened kit can be stored at 2-8℃ for 6 months. After test, the unused wells and reagents should be stored according to the table.
| Item | Specifications | Storage |
|---|---|---|
| Micro ELISA Plate(Dismountable) | 96T: 8 wells ×12 strips 48T: 8 wells ×6 strips 24T: 8 wells ×3 strips 96T*5: 5 plates, 96T |
2-8℃, 1 months |
| Reference Standard | 96T: 2 vials 48T/24T: 1 vial 96T*5: 10 vials |
2-8℃, use the reconstituted standard within 24h |
| Concentrated HRP Linked Detection Ab(100×) | 96T: 1 vial, 60 μL 48T/24T: 1 vial, 30 μL 96T*5: 5 vials, 60 μL |
2-8℃(Protect from light) |
| Reference Standard & Sample Diluent | 96T/48T/24T: 1 vial, 20 mL 96T*5: 5 vials, 20 mL |
2-8℃ |
| HRP Linked Ab Diluent | 96T/48T/24T: 1 vial, 14 mL 96T*5: 5 vials, 14 mL |
|
| Concentrated Wash Buffer(25×) | 96T/48T/24T: 1 vial, 30 mL 96T*5: 5 vials, 30 mL |
|
| Substrate Reagent | 96T/48T/24T: 1 vial, 10 mL 96T*5: 5 vials, 10 mL |
2-8℃(Protect from light) |
| Stop Solution | 96T/48T/24T: 1 vial, 10 mL 96T*5: 5 vials, 10 mL |
2-8℃ |
| Plate Sealer | 96T/48T/24T: 5 pieces 96T*5: 25 pieces |
|
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
Technical Data:
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Cortisol were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Cortisol were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 18.87 | 58.54 | 147.36 | 18.78 | 63.49 | 141.75 |
| Standard deviation | 1.28 | 3.40 | 5.56 | 0.97 | 3.58 | 5.07 |
| CV (%) | 6.79 | 5.80 | 3.77 | 5.19 | 5.64 | 3.58 |
Recovery
The recovery of Cortisol spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 84-96 | 90 |
| EDTA plasma(n=8) | 90-103 | 96 |
| Urine(n=8) | 91-109 | 99 |
Target Information
| Synonyms | 11β,17α,21-Trihydroxypregn-4-ene-3,20-dione |
| Research Area | Cell biology, Metabolism |
Assay Procedures
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1. Add 50µL each standard and sample. Immediately add 50µL of HRP linked Ab working solution. Incubate for 60 min at 37°C. |
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2. Aspirate and wash the plate for 5 times. |
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3. Add 90µL of Substrate Reagent. Incubate for about 15 min at 37°C. |
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4. Add 50µL of Stop Solution. |
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5. Read the plate at 450nm immediately. Calculation of the results |