Species:
Universal
Alternative Name:
Pregnenolone
Assay Type:
Competitive
unit:
ng/mL
Standard:
50 ng/mL
Sensitivity:
0.47 ng/mL
Detection Range:
0.78-50 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Reproductive science
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with P5. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to P5. 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 P5 in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 50 | 0.273 | |
| 25 | 0.373 | |
| 12.5 | 0.567 | |
| 6.25 | 0.828 | |
| 3.13 | 1.123 | |
| 1.56 | 1.518 | |
| 0.78 | 1.832 | |
| 0 | 2.496 |
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) | 19.69 | 42.02 | 280.9 | 20.49 | 31.45 | 349.65 |
| Standard deviation | 2.1 | 4.64 | 14.51 | 2.61 | 5.77 | 18.06 |
| C V (%) | 5.12 | 7.11 | 4.59 | 3.2 | 6.73 | 4.22 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 90-107 | 98 |
| EDTA plasma (n=8) | 87-99 | 93 |
| Cell culture media (n=8) | 82-98 | 90 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 93-102 | 85-100 | 88-102 |
| Average (%) | 101 | 100 | 93 | |
| 1:4 | Range (%) | 90-99 | 85-100 | 86-93 |
| Average (%) | 92 | 93 | 88 | |
| 1:8 | Range (%) | 95-106 | 84-98 | 91-104 |
| Average (%) | 96 | 87 | 104 | |
| 1:16 | Range (%) | 87-101 | 95-101 | 87-98 |
| Average (%) | 94 | 99 | 89 |