Species:
Universal
Alternative Name:
Phylloquinone; Phytonadione; Phytomenadione
Assay Type:
Competitive
unit:
pg/mL
Standard:
10000 pg/mL
Sensitivity:
49.3 pg/mL
Detection Range:
156.25-10000 pg/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Hematology;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 VK1. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to VK1. 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 VK1 in the samples is then determined by comparing the OD of the samples to the standard curve.
| pg/mL | OD | Corrected |
|---|---|---|
| 10000 | 0.26 | |
| 5000 | 0.36 | |
| 2500 | 0.554 | |
| 1250 | 0.815 | |
| 625 | 1.11 | |
| 312.5 | 1.505 | |
| 156.25 | 1.819 | |
| 0 | 2.483 |
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 (%) | 5.34 | 6.75 | 5.33 | 4.45 | 5.18 | 4.89 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 83-95 | 89 |
| EDTA plasma (n=8) | 95-107 | 101 |
| Cell culture media (n=8) | 82-94 | 88 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 87-96 | 87-98 | 89-102 |
| Average (%) | 89 | 89 | 99 | |
| 1:4 | Range (%) | 82-95 | 87-98 | 82-97 |
| Average (%) | 91 | 91 | 85 | |
| 1:8 | Range (%) | 92-105 | 92-101 | 88-105 |
| Average (%) | 94 | 96 | 89 | |
| 1:16 | Range (%) | 89-97 | 82-98 | 83-96 |
| Average (%) | 94 | 83 | 90 |