ELISA Kit for Vitamin D3 (VD3) 
Cholecalciferol; Toxiferol; Activated 7-Dehydrocholesterol
- UOM
- FOB US$ 557.00 US$ 796.00 US$ 3,580.00 US$ 6,763.00 US$ 55,692.00
- Quantity
Overview
Properties
- Product No.CEA920Ge
- Organism SpeciesPan-species (General) Same name, Different species.
- ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryMetabolic pathwayReproductive scienceNutrition metabolismBone metabolism
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Packages (Simulation)
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Packages (Simulation)
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Results demonstration
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Typical Standard Curve
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ISO9001: 2008, ISO13485: 2003 Registered
Recovery
Matrices listed below were spiked with certain level of Vitamin D3 (VD3) and the recovery rates were calculated by comparing the measured value to the expected amount of Vitamin D3 (VD3) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 88-95 | 92 |
| EDTA plasma(n=5) | 94-101 | 98 |
| heparin plasma(n=5) | 82-89 | 86 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vitamin D3 (VD3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vitamin D3 (VD3) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Vitamin D3 (VD3) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 87-96% | 85-101% | 81-99% | 78-94% |
| EDTA plasma(n=5) | 97-105% | 91-105% | 85-103% | 79-94% |
| heparin plasma(n=5) | 89-96% | 91-99% | 89-103% | 81-94% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
| Reagents | Quantity | Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 | Plate sealer for 96 wells | 4 |
| Standard | 2 | Standard Diluent | 1×20mL |
| Detection Reagent A | 1×120µL | Assay Diluent A | 1×12mL |
| Detection Reagent B | 1×120µL | Assay Diluent B | 1×12mL |
| TMB Substrate | 1×9mL | Stop Solution | 1×6mL |
| Wash Buffer (30 × concentrate) | 1×20mL | Instruction manual | 1 |
Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
Test principle
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Vitamin D3 (VD3) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Vitamin D3 (VD3) and unlabeled Vitamin D3 (VD3) (Standards or samples) with the pre-coated antibody specific to Vitamin D3 (VD3). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Vitamin D3 (VD3) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Vitamin D3 (VD3) in the sample.
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Citations
- Impact of Curcumin and Flaxseed Oil on Bone Turnover in Glucocorticoid-Injected Female Ratsprofile:303311214
- Protective Effect of Soybean and Mushroom against Glucocorticoid-Induced Osteoporosis in Female Rats:
- Deficiencia de vitamina D en madres y neonatos mexicanospubmed:29099117
- P38/TRHr-Dependent Regulation of TPO in Thyroid Cells Contributes to the Hypothyroidism of Triclosan-Treated RatsPubmed:29462796