Species:
Universal
Alternative Name:
Pyridoxine
Assay Type:
Competitive
unit:
ng/mL
Standard:
300 ng/mL
Sensitivity:
1.42 ng/mL
Detection Range:
4.69-300 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Nutrition metabolism;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 VB6. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to VB6. 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 VB6 in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 300 | 0.259 | |
| 150 | 0.381 | |
| 75 | 0.559 | |
| 37.5 | 0.843 | |
| 18.75 | 1.152 | |
| 9.38 | 1.52 | |
| 4.69 | 1.793 | |
| 0 | 2.541 |
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) | 5.91 | 12.61 | 84.27 | 6.15 | 9.43 | 104.9 |
| Standard deviation | 0.63 | 1.39 | 4.35 | 0.78 | 1.73 | 5.42 |
| C V (%) | 5.71 | 6.9 | 2.49 | 6.32 | 7.84 | 2.88 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 87-99 | 93 |
| EDTA plasma (n=8) | 90-104 | 97 |
| Cell culture media (n=8) | 80-97 | 88 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 85-92 | 86-100 | 87-96 |
| Average (%) | 91 | 87 | 92 | |
| 1:4 | Range (%) | 80-96 | 86-100 | 96-105 |
| Average (%) | 95 | 88 | 96 | |
| 1:8 | Range (%) | 89-101 | 93-108 | 95-103 |
| Average (%) | 95 | 102 | 101 | |
| 1:16 | Range (%) | 85-97 | 80-101 | 95-103 |
| Average (%) | 92 | 88 | 97 |