Species:
Universal
Alternative Name:
Ergocalciferol; Deltalin; Drisdol; Calcidol
Assay Type:
Competitive
unit:
ng/mL
Standard:
100 ng/mL
Sensitivity:
0.53 ng/mL
Detection Range:
1.57-100 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids.
Research:
Metabolic pathway;Neuro science;Nutrition metabolism;Bone metabolism;
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with VD2. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to VD2. 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 VD2 in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 100 | 0.278 | |
| 50 | 0.378 | |
| 25 | 0.572 | |
| 12.5 | 0.833 | |
| 6.25 | 1.128 | |
| 3.13 | 1.523 | |
| 1.56 | 1.837 | |
| 0 | 2.501 |
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) | 1.97 | 4.2 | 28.09 | 2.05 | 3.14 | 34.97 |
| Standard deviation | 0.21 | 0.46 | 1.45 | 0.26 | 0.58 | 1.81 |
| C V (%) | 4.21 | 5.32 | 2.5 | 3.47 | 7.47 | 5.62 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 92-106 | 99 |
| EDTA plasma (n=8) | 88-103 | 95 |
| Cell culture media (n=8) | 80-95 | 86 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 87-103 | 85-95 | 87-101 |
| Average (%) | 92 | 90 | 93 | |
| 1:4 | Range (%) | 97-105 | 85-95 | 88-100 |
| Average (%) | 105 | 90 | 93 | |
| 1:8 | Range (%) | 80-90 | 85-96 | 95-103 |
| Average (%) | 87 | 86 | 96 | |
| 1:16 | Range (%) | 87-96 | 93-99 | 89-97 |
| Average (%) | 94 | 95 | 94 |