ELISA Kit for 25-Hydroxyvitamin D3 (HVD3)
25-HVD3; 25HVD3; Calcifediol; Calcidiol; H-VD3; 25-Hydroxycholecalciferol; 25-Hydroxy Vitamin D3; 25(OH)D
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- FOB US$ 557.00 US$ 796.00 US$ 3,580.00 US$ 6,763.00 US$ 55,692.00
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Overview
Properties
- Product No.CEA915Ge
- Organism SpeciesPan-species (General) Same name, Different species.
- ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryReproductive scienceGenetic scienceNutrition metabolismBone metabolism
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Recovery
Matrices listed below were spiked with certain level of 25-Hydroxyvitamin D3 (HVD3) and the recovery rates were calculated by comparing the measured value to the expected amount of 25-Hydroxyvitamin D3 (HVD3) in samples.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 99-105 | 102 |
EDTA plasma(n=5) | 78-89 | 85 |
heparin plasma(n=5) | 93-105 | 98 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 25-Hydroxyvitamin D3 (HVD3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 25-Hydroxyvitamin D3 (HVD3) 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 25-Hydroxyvitamin D3 (HVD3) 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) | 82-91% | 81-104% | 87-99% | 91-101% |
EDTA plasma(n=5) | 80-95% | 83-102% | 79-99% | 97-104% |
heparin plasma(n=5) | 86-104% | 90-97% | 93-105% | 78-98% |
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 HVD3 has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled HVD3 analogues and unlabeled antigen (Standards or samples) with the pre-coated antibody. 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 HVD3 in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of HVD3 in the sample.
Giveaways
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Citations
- Decreased serum LL-37 and vitamin D3 levels in atopic dermatitis: relationship between IL-31 and oncostatin MWiley: source
- 成人哮喘血清维生素D 水平与肺功能相关因素分析Jwk_Tjyy: Cn
- Comparative effects of brown and golden flaxseeds on body composition, inflammation and bone remodelling biomarkers in perimenopausal overweight womenS1756464617301561
- Chronic maternal calcium and 25-hydroxyvitamin D deficiency in Wistar rats programs abnormal hepatic gene expression leading to hepatic steatosis in female offspring.pubmed:28219837
- Effect of Calcitriol on FGF23 Level in Healthy Adults and its Dependence on Phosphate Levelpubmed:28064234
- Serum 25-hydroxyvitamin D inversely associated with blood eosinophils in patients with persistent allergic rhinitis.pubmed:29094019
- Effects of Vitamin D Restricted Diet Administered during Perinatal and Postnatal Periods on the Penis of Wistar RatsPubmed:29850540
- Evaluation of FGF‐23 and 25(OH)D3 levels in peri‐implant sulcus fluid in peri‐implant health and diseasesPubmed: 31407857
- Circulating Osteocyte‐Related Biomarkers (vitamin D, sclerostin, dickkopf-1), hepcidin, and oxidative stress markers in early breast cancer: their impact in …Pubmed: 33065276
- The Effects of Follicular Fluid 25 (OH) D Concentration on Intrafollicular Estradiol Level, Oocyte Quality, and Fertilization Rate in Women Who Underwent IVF Program