Human Vitamin D BP Quantikine ELISA Kit

DVDBP0 has been discontinued and is replaced by DVDBP0B.
(17 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours
  • Specificity
    Natural human Vitamin D Binding Protein
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Human Vitamin D Binding Protein immunoassay is a 3.5 hour solid phase ELISA designed to measure Vitamin D Binding Protein in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains natural human Vitamin D Binding Protein Standard. This kit can be used to determine mass values for naturally occurring human Vitamin D Binding Protein.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean3312818052.9105164
Standard Deviation1.887.4211.22.76.312.1
CV%5.75.86.25.167.4

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean10.931.763.710.130.861.9
Standard Deviation0.2440.4231.360.6610.6641.31
CV%2.21.32.16.52.22.1

Recovery

The recovery of Vitamin D Binding Protein spiked to three different levels throughout the range of the assay was evaluated.

Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of Vitamin D Binding Protein were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Vitamin D BP
Vitamin D binding protein (VitD BP), also known as DBP and Gc-globulin, is a 58 kDa glycoprotein that circulates at high concentration in the serum and serves as a carrier protein for vitamin D. The transport of vitamin D by VitD BP is important for the function of a wide variety of tissues. VitD BP binds both the 25(OH) and the hormonally active 1,25(OH)2 forms of vitamin D. VitD BP is primarily expressed in hepatocytes and to a lesser extent in the kidney. It delivers vitamin D into cells by Megalin-mediated endocytosis. A selectively deglycosylated form of VitD BP known as macrophage activating factor (MAF) blocks the angiogenic effects of FGF basic, VEGF, and Angiopoietin 2. VitD BP enhances the chemotaxis of monocytes and neutrophils to the activated complement component C5a or C5a des Arg (a C-terminally processed form of C5a). The chemotactic cofactor property of VitD BP is eliminated by binding to 1,25(OH)2 vitamin D, but it is not altered by binding to 25(OH) vitamin D or actin.
    • Long Name
      Vitamin D Binding Protein
    • Entrez Gene IDs
      2638 (Human); 14473 (Mouse); 24384 (Rat);
    • Alternate Names
      DBP; DBP/GC; DBP-MAF; DBPVDBG; EC 6.3.1.5; Gc; gc-globulin; Gc-MAF; GRD3; group-specific component (vitamin D binding protein); Group-specific component; hDBP; VDB; VDBG; VDBP; Vitamin D BP; vitamin D-binding alpha-globulin; vitamin D-binding protein;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    17 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Bioavailable 25(OH)D but Not Total 25(OH)D Is an Independent Determinant for Bone Mineral Density in Chinese Postmenopausal Women
      Authors: C Li, P Chen, X Duan, J Wang, B Shu, X Li, Q Ba, J Li, Y Wang, H Wang
      EBioMedicine, 2017;15(0):184-192.
      Species: Human
      Sample Type: Plasma
    2. Genetically Low Vitamin D Levels, Bone Mineral Density, and Bone Metabolism Markers: a Mendelian Randomisation Study
      Sci Rep, 2016;6(0):33202.
      Species: Human
      Sample Type: Serum
    3. Pharmacokinetics and effects of demographic factors on blood 25(OH)D3 levels after a single orally administered high dose of vitamin D3
      Acta Pharmacol Sin, 2016;0(0):.
      Species: Human
      Sample Type: Plasma
    4. Urinary Vitamin D Binding Protein and KIM-1 Are Potent New Biomarkers of Major Adverse Renal Events in Patients Undergoing Coronary Angiography.
      Authors: Chaykovska L, Heunisch F, von Einem G, Alter M, Hocher C, Tsuprykov O, Dschietzig T, Kretschmer A, Hocher B
      PLoS ONE, 2016;11(1):e0145723.
      Species: Human
      Sample Type: Plasma
    5. Serum vitamin D, vitamin D binding protein, and risk of colorectal cancer.
      Authors: Anic G, Weinstein S, Mondul A, Mannisto S, Albanes D
      PLoS ONE, 2014;9(7):e102966.
      Species: Human
      Sample Type: Serum
    6. Vitamin D-binding protein, circulating vitamin D and risk of renal cell carcinoma.
      Authors: Mondul A, Weinstein S, Moy K, Mannisto S, Albanes D
      Int J Cancer, 2014;134(11):2699-706.
      Species: Human
      Sample Type: Serum
    7. Associations between vascular health indices and serum total, free and bioavailable 25-hydroxyvitamin D in adolescents.
      Authors: Ashraf A, Alvarez J, Dudenbostel T, Calhoun D, Griffin R, Wang X, Hanks L, Gower B
      PLoS ONE, 2014;9(12):e114689.
      Species: Human
      Sample Type: Serum
    8. Serum vitamin D levels are positively associated with varicella zoster immunity in chronic dialysis patients.
      Authors: Chao C, Lee S, Yang W, Yen C, Chiang C, Huang J, Hung K
      Sci Rep, 2014;4(0):7371.
      Species: Human
      Sample Type: Serum
    9. A bone substitute with high affinity for vitamin D-binding protein--relationship with niche of osteoclasts.
      Authors: Ikeda T, Kasai M, Tatsukawa E, Kamitakahara M, Shibata Y, Yokoi T, Nemoto T, Ioku K
      J Cell Mol Med, 2014;18(1):170-80.
      Species: Human
      Sample Type: Serum
    10. Circulating 25-hydroxyvitamin D, vitamin D-binding protein and risk of prostate cancer.
      Authors: Weinstein S, Mondul A, Kopp W, Rager H, Virtamo J, Albanes D
      Int J Cancer, 2013;132(12):2940-7.
      Species: Human
      Sample Type: Serum
    11. Influence of vitamin D binding protein on the association between circulating vitamin D and risk of bladder cancer.
      Br. J. Cancer, 2012;107(9):1589-94.
      Species: Human
      Sample Type: Serum
    12. Accuracy of 6 routine 25-hydroxyvitamin D assays: influence of vitamin D binding protein concentration.
      Authors: Heijboer AC, Blankenstein MA
      Clin. Chem., 2012;58(3):543-8.
      Species: Human
      Sample Type: Plasma
    13. Bioavailable vitamin D is more tightly linked to mineral metabolism than total vitamin D in incident hemodialysis patients.
      Authors: Bhan I, Powe CE, Berg AH
      Kidney Int., 2012;82(1):84-9.
      Species: Human
      Sample Type: Serum
    14. Vitamin D in a northern Canadian first nation population: dietary intake, serum concentrations and functional gene polymorphisms.
      Authors: Larcombe L, Mookherjee N, Slater J, Slivinski C, Singer M, Whaley C, Denechezhe L, Matyas S, Turner-Brannen E, Nickerson P, Orr P
      PLoS ONE, 2012;7(11):e49872.
      Species: Human
      Sample Type: Serum
    15. Oral cholecalciferol decreases albuminuria and urinary TGF-beta1 in patients with type 2 diabetic nephropathy on established renin-angiotensin-aldosterone system inhibition.
      Authors: Kim MJ, Frankel AH, Donaldson M, Darch SJ, Pusey CD, Hill PD, Mayr M, Tam FW
      Kidney Int., 2011;80(8):851-60.
      Species: Human
      Sample Type: Urine
    16. Differences in bone metabolism between singleton pregnancy and twin pregnancy.
      Authors: Nakayama S, Yasui T, Suto M, Sato M, Kaji T, Uemura H, Maeda K, Irahara M
      Bone, 2011;49(3):513-9.
      Species: Human
      Sample Type: Serum
    17. Vitamin D supplementation during pregnancy: double-blind, randomized clinical trial of safety and effectiveness.
      Authors: Hollis BW, Johnson D, Hulsey TC, Ebeling M, Wagner CL
      J. Bone Miner. Res., 2011;26(10):2341-57.
      Species: Human
      Sample Type: Serum
    Expand to show all 17 Citations
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    Supplemental ELISA Products
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    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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      DVDBP0 has been replaced by
      DVDBP0B

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