Species:
Universal
Alternative Name:
17-Ketosteroid; 17-KS; 3a-Hydroxy-17-Androstanone
Assay Type:
Competitive
unit:
pg/mL
Standard:
90000 pg/mL
Sensitivity:
144 pg/mL
Detection Range:
1406.25-90000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Endocrinology;Reproductive science;Hormone metabolism;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with ADT. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to ADT. 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 ADT in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 90000 | 0.434 | |
| 45000 | 0.55 | |
| 22500 | 0.746 | |
| 11250 | 1.031 | |
| 5625 | 1.369 | |
| 2812.5 | 1.684 | |
| 1406.25 | 1.92 | |
| 0 | 2.439 |
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) | 1771.65 | 3781.51 | 25280.9 | 1844.26 | 2830.19 | 31468.53 |
| Standard deviation | 188.78 | 417.2 | 1305.83 | 234.98 | 519.31 | 1625.44 |
| C V (%) | 5.05 | 6.64 | 4.15 | 3.8 | 5.33 | 3.56 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 87-95 | 91 |
| EDTA plasma (n=8) | 81-95 | 88 |
| Cell culture media (n=8) | 83-97 | 90 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-94 | 85-103 | 87-98 |
| Average (%) | 87 | 101 | 91 | |
| 1:4 | Range (%) | 90-99 | 85-103 | 80-92 |
| Average (%) | 92 | 87 | 85 | |
| 1:8 | Range (%) | 82-101 | 83-96 | 96-105 |
| Average (%) | 82 | 94 | 104 | |
| 1:16 | Range (%) | 91-102 | 95-102 | 86-97 |
| Average (%) | 96 | 97 | 92 |