Test Principle
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Cor. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Cor. 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 Cor in the samples is then determined by comparing the OD of the samples to the standard curve.
|
Product Name |
Cor(Cortisol) ELISA Kit |
|
Alternative Names |
Hydrocortisone |
|
Assay Type |
Competitive Inhibition |
|
Standard |
200 ng/mL |
|
Sample Type |
serum, plasma, urine and other biological fluids |
|
Assay Length |
2h |
|
Research Area |
Metabolic pathway, Endocrinology, Hormone metabolism |
Standard Curve
|
Concentration (pg/mL) |
OD |
Corrected OD |
|
200.00 |
0.197 |
- |
|
100.00 |
0.397 |
- |
|
50.00 |
0.451 |
- |
|
25.00 |
0.897 |
- |
|
12.50 |
1.236 |
- |
|
6.25 |
1.457 |
- |
|
3.13 |
1.877 |
- |
|
0.00 |
2.232 |
- |
Precision
-
- Intra-assay Precision (Precision within an assay)：CV%<8%
- Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
- Inter-assay Precision (Precision between assays)：CV%<10%
- Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant Cor and the recovery rates were calculated by comparing the measured value to the expected amount of Cor in samples.
|
Matrix |
Recovery range |
Average |
|
serum(n=5) |
87-99% |
93% |
|
EDTA plasma(n=5) |
93-107% |
100% |
|
Heparin plasma(n=5) |
85-99% |
93% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cor and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
|
Matrix |
1:2 |
1:4 |
1:8 |
1:16 |
|
serum(n=5) |
93-102% |
93-106% |
82-101% |
91-103% |
|
EDTA plasma(n=5) |
83-95% |
88-102% |
85-97% |
80-92% |
|
Heparin plasma(n=5) |
82-93% |
95-104% |
93-102% |
79-94% |
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