Species:
Universal
Alternative Name:
VA
Assay Type:
Competitive
unit:
ng/mL
Standard:
1000 ng/mL
Sensitivity:
9.38 ng/mL
Detection Range:
15.63-1000 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids
Research:
Metabolic pathway;Endocrinology;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 VA. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to VA. 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 VA in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 1000 | 0.264 | |
| 500 | 0.364 | |
| 250 | 0.558 | |
| 125 | 0.819 | |
| 62.5 | 1.114 | |
| 31.25 | 1.509 | |
| 15.63 | 1.823 | |
| 0 | 2.487 |
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 (%) | 5.46 | 7.62 | 5.37 | 4.02 | 5.74 | 2.28 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 89-103 | 96 |
| EDTA plasma (n=8) | 83-97 | 90 |
| Cell culture media (n=8) | 86-99 | 92 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 86-93 | 92-101 | 80-96 |
| Average (%) | 87 | 94 | 92 | |
| 1:4 | Range (%) | 93-101 | 92-101 | 95-103 |
| Average (%) | 98 | 100 | 101 | |
| 1:8 | Range (%) | 85-98 | 87-98 | 80-96 |
| Average (%) | 91 | 92 | 95 | |
| 1:16 | Range (%) | 90-101 | 85-92 | 93-106 |
| Average (%) | 93 | 87 | 99 |