VD(Vitamin D) ELISA Kit
SKU: E-EL-0012
VD(Vitamin D) ELISA Kit
| Detection Range | 6.25-400 ng/mL |
| Sensitivity | 3.75 ng/mL |
Product Details
Properties
| Assay Type | Competitive-ELISA |
| size | 96T |
| Assay Time | 2.5h |
| Detection Method | Colormetric |
| Detection Range | 6.25-400 ng/mL |
| Sensitivity | 3.75 ng/mL |
| Sample Volume | 50μL |
| Sample Type | serum, plasma and other biological fluids |
| Specificity | This kit recognizes Universal VD in samples.No significant cross-reactivity or interference between Universal VD and analogues was observed |
| Precision | Both intra-CV and inter-CV are < 10%. |
| Recovery | 80%-120% |
| Introduction | This ELISA kit applies to the in vitro quantitative determination of VD concentrations in serum, plasma and other biological fluids. |
| Applications | ELISA |
| Storage | 2-8℃/-20℃ |
Test Principle
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal VD. During the reaction, Universal VD in the sample or standard competes with a fixed amount of Universal VD on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal VD. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal VD in tested samples can be calculated by comparing the OD of the samples to the standard curve.
Kit Components and Storage
An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 96T: 8 wells ×12 strips | -20℃, 6 months |
| 48T: 8 wells ×6 strips | ||
| Reference Standard | 96T: 2 vials | |
| 48T: 1 vial | ||
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL | |
| 48T: 1 vial, 60 μL | ||
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL | -20℃(Protect from light), 6 months |
| 48T: 1 vial, 60 μL | ||
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 2-8℃ (Protect from light) |
| Stop Solution | 1 vial, 10 mL | 2-8℃ |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy |
Technical Data:
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and
high level VD were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and
high level VD 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) | 21.62 | 53.66 | 157.91 | 21.41 | 57.3 | 152.81 |
| Standard deviation | 1.28 | 2.7 | 5.98 | 1.47 | 2.35 | 8.34 |
| CV (%) | 5.92 | 5.03 | 3.79 | 6.87 | 4.1 | 5.46 |
Recovery
The recovery of VD 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) | 96-112 | 102 |
| EDTA plasma(n=8) | 88-99 | 93 |
| Cell culture media(n=8) | 85-97 | 91 |
Target Information
| Research Area | Metabolism |
Assay Procedures
 |
1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C |
 |
2. Aspirate and wash the plate for 3 times |
 |
3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times |
 |
4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C |
 |
5. Add 50μL Stop Solution |
 |
6. Read the plate at 450nm immediately. Calculation of the results |