Multiplex Assay Kit for Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay)

NR1I1; Nuclear Receptor Subfamily 1 Group I Member 1; Calcitriol Receptor; 1,25-dihydroxyvitamin D3 receptor

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-103 99
EDTA plasma(n=5) 80-104 83
heparin plasma(n=5) 83-102 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Vitamin D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 D Receptor (VDR) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 88-103% 86-103% 87-101% 90-104%
EDTA plasma(n=5) 91-99% 83-103% 85-94% 81-92%
heparin plasma(n=5) 89-96% 92-101% 94-103% 98-105%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:VDR) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
6 The vitamin D receptor: A therapeutic target for the treatment of breast cancer? Cgi: Content
endocrine-related cancer Vitamin D receptor as a target for breast cancer therapy. pubmed:28213567
Journal of Advanced Research Effect of exercise on serum vitamin D and tissue vitamin Dreceptors in experimentally induced type 2 Diabetes Mellitus. pubmed:27504197
European Journal of Applied Physiology Vitamin D supplementation attenuates oxidative stress in paraspinal skeletal muscles in patients with low back pain Pubmed:29143122
BBA- Molecular Basis of Disease Enhanced remedial effects for vitamin D3 and calcium co-supplementation against pre-existing lead nephrotoxicity in mice: The roles of renal calcium homeostatic … Pubmed: 30553018
OPEN ACCESS MACEDONIAN JOURNAL OF MEDICAL SCIENCES Seluang Fish (Rasbora Spp.) Oil Decreases Inflammatory Cytokines Via Increasing Vitamin D Level in Systemic Lupus Erythematosus
Journal of Steroid Biochemistry and Molecular Biology Status of vitamin D and the associated host factors in pulmonary tuberculosis patients and their household contacts: a cross sectional study Pubmed: 31255688
Scientific Reports Association of Fok1 VDR polymorphism with Vitamin D and its associated molecules in pulmonary tuberculosis patients and their household contacts Pubmed: 31649297
Research square Vitamin D Status in Dupuytren's Disease: Association with Clinical Status and Vitamin D Receptor Expression
Betok Fish (Anabas testudineus) Oil Decreases Inflammatory Cytokine through Increasing Vitamin D Level in Rats-induced Systemic Lupus Erythematosus
iium medical journal malaysia Positive Correlation between Monocyte-to-Lymphocyte Ratio and C-Reactive Protein in Vitamin D Deficient Preterm Infants with Respiratory Distress Syndrome
Research Square Identification of SNP of VDR and VDBP gene and its Dysregulated pathway through VDR-VDBP interaction network analysis in Vitamin D deficient Infertile …
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