ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3) Pan-species (General) Competition ELISA

DH-VD3; Calcitriol; Rocaltrol; Calcijex; 1,25-(OH)2D3; 1,25-Dihydroxycholecalciferol; 1,25-Dihydroxy Vitamin D3

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  • ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3) Packages (Simulation)
  • ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3) Packages (Simulation)
  • ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3) Results demonstration
  • CEA467Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of 1,25-Dihydroxyvitamin D3 (DHVD3) and the recovery rates were calculated by comparing the measured value to the expected amount of 1,25-Dihydroxyvitamin D3 (DHVD3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-104 101
EDTA plasma(n=5) 83-95 87
heparin plasma(n=5) 86-98 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 1,25-Dihydroxyvitamin D3 (DHVD3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 1,25-Dihydroxyvitamin D3 (DHVD3) 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 1,25-Dihydroxyvitamin D3 (DHVD3) 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) 93-101% 81-92% 88-98% 89-96%
EDTA plasma(n=5) 87-102% 92-101% 92-99% 92-101%
heparin plasma(n=5) 78-89% 93-101% 78-103% 96-103%

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.

ELISA Kit for 1,25-Dihydroxyvitamin D3 (DHVD3)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to DHVD3 has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled DHVD3 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 DHVD3 in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of DHVD3 in the sample.

Citations

  • Selective Cytotoxic Action and DNA Damage by Calcitriol-Cu(II) Interaction: Putative Mechanism of Cancer PreventionPubMed: PMC3785422
  • Calcium-deficiency assessment and biomarker identification by an integrated urinary metabonomics analysisPubMed: PMC3652781
  • Treatment of established left ventricular hypertrophy with fibroblast growth factor receptor blockade in an animal model of CKDPubmed:24875663
  • Vitamin D Prevents Podocyte Injury via Regulation of Macrophage M1/M2 Phenotype in Diabetic Nephropathy RatsPubmed:25188527
  • Calcitriol?induced?redox?imbalance and DNA breakage in cells sharing a common metabolic feature of malignancies: Interaction with cellular copper (II) ions leads to the production of reactive oxygen species.Pubmed:25547436
  • 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:
  • Calcitriol–copper interaction leads to non enzymatic, reactive oxygen species mediated DNA breakage and modulation of cellular redox scavengers in hepatocellular …article:10.1007
  • Effect of Calcitriol on FGF23 Level in Healthy Adults and its Dependence on Phosphate Levelpubmed:28064234
  • Glucocorticoid exposure induces preeclampsia via dampening 1,25-dihydroxyvitamin D3pubmed:29093562
  • Combined effect of pulsed electromagnetic fields and narrowband ultraviolet B on bone metabolism in glucocorticoid-treated rats
  • Dobesity: relationship between vitamin d deficiency, obesity and sclerostin as a novel biomarker of bone metabolism
  • Alleviating Effect of α-Lipoic Acid and Magnesium on Cadmium-Induced Inflammatory Processes, Oxidative Stress and Bone Metabolism Disorders in Wistar RatsPubmed: 31739465
  • Sodium dehydroacetate induces cardiovascular toxicity associated with Ca2+ imbalance in zebrafish
  • Metabolic activities affect femur and lumbar vertebrae remodeling, and anti-resorptive risedronate disturbs femoral cortical bone remodeling33436949
  • SWIM domain protein ZSWIM4 is required for JAK2 inhibition resistance in breast cancer34102191
  • Effect of Dapagliflozin and Magnesium Supplementation on Renal Magnesium Handling and Magnesium Homeostasis in Metabolic Syndrome34836340

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