Species:
Universal
Alternative Name:
Riboflavin; Riboflavine
Assay Type:
Competitive
unit:
ng/mL
Standard:
600 ng/mL
Sensitivity:
2.55 ng/mL
Detection Range:
9.38-600 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates 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 VB2. Standards or samples are added to the appropriate micro ELISA plate wells then with a biotin-conjugated antibody specific to VB2. 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 VB2 in the samples is then determined by comparing the OD of the samples to the standard curve.
| ng/mL | OD | Corrected |
|---|---|---|
| 600 | 0.197 | |
| 300 | 0.375 | |
| 150 | 0.614 | |
| 75 | 0.859 | |
| 37.5 | 1.184 | |
| 18.75 | 1.543 | |
| 9.38 | 1.788 | |
| 0 | 2.548 |
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) | 11.81 | 25.21 | 168.54 | 12.3 | 18.87 | 209.79 |
| Standard deviation | 1.26 | 2.78 | 8.71 | 1.57 | 3.46 | 10.84 |
| C V (%) | 4.71 | 4.56 | 3.32 | 4.58 | 7.21 | 3.75 |
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-94 | 87 |
| EDTA plasma (n=8) | 95-107 | 101 |
| Cell culture media (n=8) | 82-95 | 88 |
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 88-102 | 90-99 | 82-97 |
| Average (%) | 93 | 97 | 86 | |
| 1:4 | Range (%) | 93-105 | 90-99 | 88-103 |
| Average (%) | 101 | 91 | 103 | |
| 1:8 | Range (%) | 91-99 | 95-106 | 83-94 |
| Average (%) | 98 | 106 | 84 | |
| 1:16 | Range (%) | 83-96 | 87-101 | 92-105 |
| Average (%) | 91 | 100 | 100 |