ELISA Kit for 25-Hydroxyvitamin D3 (HVD3)

25-HVD3; 25HVD3; Calcifediol; Calcidiol; H-VD3; 25-Hydroxycholecalciferol; 25-Hydroxy Vitamin D3; 25(OH)D

Specificity

This assay has high sensitivity and excellent specificity for detection of 25-Hydroxyvitamin D3 (HVD3).
No significant cross-reactivity or interference between 25-Hydroxyvitamin D3 (HVD3) and analogues was observed.

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.

GIVEAWAYS

INCREMENT SERVICES

Magazine Citations
Allergy & Clinical Immunology Decreased serum LL-37 and vitamin D3 levels in atopic dermatitis: relationship between IL-31 and oncostatin M Wiley: source
Tianjin Med J 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
Journal of Functional Foods Comparative effects of brown and golden flaxseeds on body composition, inflammation and bone remodelling biomarkers in perimenopausal overweight women S1756464617301561
J Nutr Biochem. 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
 in vivo Effect of Calcitriol on FGF23 Level in Healthy Adults and its Dependence on Phosphate Level pubmed:28064234
Asia pacific allergy Serum 25-hydroxyvitamin D inversely associated with blood eosinophils in patients with persistent allergic rhinitis. pubmed:29094019
Biomed Research International  Effects of Vitamin D Restricted Diet Administered during Perinatal and Postnatal Periods on the Penis of Wistar Rats Pubmed:29850540
Clin Implant Dent Relat Res. Evaluation of FGF‐23 and 25(OH)D3 levels in peri‐implant sulcus fluid in peri‐implant health and diseases Pubmed: 31407857
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY Circulating Osteocyte‐Related Biomarkers (vitamin D, sclerostin, dickkopf-1), hepcidin, and oxidative stress markers in early breast cancer: their impact in … Pubmed: 33065276
The Journal of Obstetrics and Gynecology of India The Effects of Follicular Fluid 25 (OH) D Concentration on Intrafollicular Estradiol Level, Oocyte Quality, and Fertilization Rate in Women Who Underwent IVF Program
Catalog No. Related products for research use of Pan-species (General) Organism species Applications (RESEARCH USE ONLY!)
CPA915Ge11 BSA Conjugated 25-Hydroxyvitamin D3 (HVD3) Immunogen; SDS-PAGE; WB.
CPA915Ge21 OVA Conjugated 25-Hydroxyvitamin D3 (HVD3) Immunogen; SDS-PAGE; WB.
PAA915Ge01 Polyclonal Antibody to 25-Hydroxyvitamin D3 (HVD3) WB; IHC; ICC; IP.
LAA915Ge71 Biotin-Linked Polyclonal Antibody to 25-Hydroxyvitamin D3 (HVD3) WB; IHC; ICC.
MAA915Ge22 Monoclonal Antibody to 25-Hydroxyvitamin D3 (HVD3) ELISA, CLIA. / IHC-Fr, ICC, IP (predicted).
MAA915Ge21 Monoclonal Antibody to 25-Hydroxyvitamin D3 (HVD3) ELISA, CLIA. / IHC-Fr, ICC, IP (predicted).
CEA915Ge ELISA Kit for 25-Hydroxyvitamin D3 (HVD3) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA915Ge Multiplex Assay Kit for 25-Hydroxyvitamin D3 (HVD3) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.
KSA915Ge11 ELISA Kit DIY Materials for 25-Hydroxyvitamin D3 (HVD3) Main materials for "Do It (ELISA Kit) Yourself".