Universal ELISA Kits
VD3(Vitamin D3) ELISA Kit
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Product No
FY-EU1114Sample Type
Serum, Plasma and Other biological fluidsAssay Time
1.5HPurchase details
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Product Details
Product Name in English:
VD3(Vitamin D3) ELISA Kit
Species:
Universal
Alternative Name:
Cholecalciferol; Toxiferol; Activated 7-Dehydrocholesterol
Assay Type:
Competitive
Standard:
400 ng/mL
Sensitivity:
2.14 ng/mL
Detection Range:
6.25-400 ng/mL
Assay Time:
1.5H
Sample Type:
Serum, Plasma and Other biological fluids
Research:
Metabolic pathway;Reproductive science;Nutrition metabolism;Bone metabolism;
Test Principle:
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with VD3 protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to VD3. 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 stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of VD3 in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration | OD1 | OD2 | Average OD | Corrected |
|---|---|---|---|---|
| 400 | 0.258 | 0.258 | 0.258 | |
| 200 | 0.421 | 0.421 | 0.421 | |
| 100 | 0.661 | 0.664 | 0.663 | |
| 50 | 0.978 | 0.988 | 0.983 | |
| 25 | 1.271 | 1.275 | 1.273 | |
| 12.5 | 1.591 | 1.592 | 1.592 | |
| 6.25 | 1.861 | 1.863 | 1.862 | |
| 0 | 2.683 | 2.704 | 2.694 |
Linear
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 86-93 | 93-102 | 91-104 |
| Average (%) | 91 | 98 | 92 | |
| 1:4 | Range (%) | 91-105 | 93-102 | 89-97 |
| Average (%) | 96 | 101 | 93 | |
| 1:8 | Range (%) | 87-98 | 93-106 | 93-102 |
| Average (%) | 90 | 102 | 97 | |
| 1:16 | Range (%) | 89-94 | 87-101 | 81-96 |
| Average (%) | 94 | 90 | 88 |
Precision
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.
| Item | Intra-assay Precision | Intra-assay Precision |
|---|---|---|
| Sample | 3 | 3 |
| Replicate | 9 | 18 |
| CV(%) | 5 | 8 |
Rate of recovery
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 92-105 | 98 |
| EDTA plasma (n=8) | 84-96 | 90 |
| Cell culture media (n=8) | 80-95 | 87 |
Assay Procedures
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