Species:
Universal
Alternative Name:
4-Androstenedione; 4-Androstene-3,17-Dione
Assay Type:
Competitive
unit:
ng/mL
Standard:
10 ng/mL
Sensitivity:
0.096 ng/mL
Detection Range:
0.16-10 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates 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 ASD. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to ASD. 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 ASD in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 10 | 0.279 | |
| 5 | 0.379 | |
| 2.5 | 0.573 | |
| 1.25 | 0.834 | |
| 0.63 | 1.129 | |
| 0.31 | 1.524 | |
| 0.16 | 1.838 | |
| 0 | 2.502 |
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) | 196.85 | 420.17 | 2808.99 | 204.92 | 314.47 | 3496.5 |
| Standard deviation | 20.98 | 46.36 | 145.09 | 26.11 | 57.7 | 180.6 |
| C V (%) | 3.23 | 7.2 | 3.34 | 3.75 | 4.22 | 5.02 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-90 | 84 |
| EDTA plasma (n=8) | 85-97 | 91 |
| Cell culture media (n=8) | 82-96 | 89 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-92 | 88-101 | 96-103 |
| Average (%) | 89 | 98 | 102 | |
| 1:4 | Range (%) | 90-96 | 88-101 | 85-94 |
| Average (%) | 91 | 99 | 90 | |
| 1:8 | Range (%) | 80-102 | 86-92 | 87-103 |
| Average (%) | 83 | 92 | 88 | |
| 1:16 | Range (%) | 83-99 | 83-98 | 81-95 |
| Average (%) | 93 | 94 | 82 |