Test Principle
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 11-KT. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 11-KT. 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 11-KT in the samples is then determined by comparing the OD of the samples to the standard curve.
| Product Name | 11-KT(11-keto Testosterone) ELISA Kit |
| Alternative Names | 11-keto Testosterone |
| Assay Type | Competitive Inhibition |
| Standard | 10000 pg/mL |
| Sample Type | serum, plasma and other biological fluids |
| Assay Length | 2h |
| Research Area | Endocrinology, Reproductive Science, Hormone Metabolism |
Standard Curve
| Concentration (pg/mL) | OD | Corrected OD |
| 10000.00 | 0.191 | - |
| 5000.00 | 0.397 | - |
| 2500.00 | 0.715 | - |
| 1250.00 | 0.823 | - |
| 625.00 | 1.257 | - |
| 312.50 | 1.572 | - |
| 156.25 | 1.769 | - |
| 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 11-KT and the recovery rates were calculated by comparing the measured value to the expected amount of 11-KT in samples.
| Matrix | Recovery range | Average |
| serum(n=5) | 90-105% | 97% |
| EDTA plasma(n=5) | 86-99% | 92% |
| Heparin plasma(n=5) | 80-95% | 86% |
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 11-KT 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) | 79-94% | 87-96% | 87-101% | 82-90% |
| EDTA plasma(n=5) | 87-95% | 78-91% | 97-103% | 95-101% |
| Heparin plasma(n=5) | 85-93% | 88-95% | 96-102% | 83-97% |
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