Species:
Universal
Alternative Name:
Cortisol
Assay Type:
Competitive
unit:
ng/mL
Standard:
20ng/mL
Sensitivity:
0.19 ng/mL
Detection Range:
0.31-20ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Other biological fluids,Sample volume: 50μL
Research:
Cardiovascular;Signalingtransduction;Neuroscience;Cancer;Metabolism
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Cortisol. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to Cortisol. Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Cortisol in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 20 | 0.279 | |
| 10 | 0.445 | |
| 5 | 0.782 | |
| 2.5 | 1.165 | |
| 1.25 | 1.539 | |
| 0.63 | 1.881 | |
| 0.31 | 2.123 | |
| 0 | 2.46 |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean(ng/mL) | 3.94 | 8.4 | 6.18 | 4.1 | 6.29 | 9.93 |
| Standard deviation | 0.42 | 0.93 | 2.9 | 0.52 | 1.15 | 3.61 |
| C V (%) | 6.23 | 6.05 | 3.54 | 4.11 | 6.96 | 5.33 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-92 | 86 |
| EDTA plasma (n=8) | 80-95 | 86 |
| Cell culture media (n=8) | 80-96 | 88 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-92 | 88-101 | 87-96 |
| Average (%) | 90 | 92 | 89 | |
| 1:4 | Range (%) | 89-96 | 85-93 | 92-105 |
| Average (%) | 91 | 86 | 97 | |
| 1:8 | Range (%) | 80-98 | 89-97 | 89-102 |
| Average (%) | 85 | 93 | 92 | |
| 1:16 | Range (%) | 81-92 | 85-94 | 92-101 |
| Average (%) | 85 | 90 | 94 |