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Multiplex Assay Kit for Vitamin D Binding Protein (DBP) ,etc. by FLIA (Flow Luminescence Immunoassay)
GC; VDBG; VDBP; Group-Specific Component; Gc-globulin
(Note: Up to 8-plex in one testing reaction)
- Product No.LMB810Ra
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- Sample TypeSerum, plasma and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length3.5h
- Detection Range0.2-200ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.067 ng/mL.
- DownloadInstruction Manual
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
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Result
For more details, please contact local distributors!
Specificity
This assay has high sensitivity and excellent specificity for detection of Vitamin D Binding Protein (DBP) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Vitamin D Binding Protein (DBP) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Vitamin D Binding Protein (DBP) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Vitamin D Binding Protein (DBP) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 78-94 | 82 |
| EDTA plasma(n=5) | 80-101 | 89 |
| heparin plasma(n=5) | 87-99 | 94 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Vitamin D Binding Protein (DBP) ,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 Binding Protein (DBP) ,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 Binding Protein (DBP) ,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) | 80-91% | 88-101% | 84-95% | 93-102% |
| EDTA plasma(n=5) | 92-101% | 78-101% | 82-97% | 98-105% |
| heparin plasma(n=5) | 79-101% | 78-105% | 87-101% | 96-104% |
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#:DBP) | 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 |
| Proteome Science | Isotope Coded Protein Labeling analysis of plasma specimens from acute severe dengue fever patients PubMed: 23101585 |
| Toxins (Basel). | Increased Circulating Levels of Vitamin D Binding Protein in MS Patients Pubmed:25590278 |
| Endocrine | Vitamin D-binding protein and free vitamin D concentrations in acromegaly PubMed: 26547217 |
| Proteomics Clinical Applications | Comparative mass spectrometric and immunoassay‐based proteome analysis in serum of Duchenne muscular dystrophy patients Pubmed:26680509 |
| PLoS One. | 25-Hydroxivitamin D Serum Concentration, Not Free and Bioavailable Vitamin D, Is Associated with Disease Activity in Systemic Lupus Erythematosus Patients. pubmed:28085957 |
| PROTEOMICS - Clinical Applications | Comparative mass spectrometric and immunoassay-based proteome analysis in serum ofDuchenne muscular dystrophy patients. pubmed:26680509 |
| Plos One | 25-Hydroxivitamin D serum concentration, not free and bioavailable vitamin D, is associated with disease activity in systemic lupus erythematosus patients 10.1371/journal.pone.0170323 |
| Nutrients | Gene Expression of Sirtuin-1 and Endogenous Secretory Receptor for Advanced Glycation End Products in Healthy and Slightly Overweight Subjects after … Pubmed:30037068 |
| 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 |
| J Clin Res Pediatr Endocrinol | Is Bioavailable Vitamin D Better Than Total Vitamin D to Evaluate Vitamin D Status in Obese Children? 34013709 |
| PLoS One | Leukocyte telomere length as a compensatory mechanism in vitamin D metabolism Pubmed:35202418 |