• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Range

  187.00 - 3,000 pg/mL

• Sufficient Materials

  For five 96-well plates*

• Specificity

  Please see the

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Vitamin D Binding Protein (Vitamin D BP). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, saliva, urine, and human milk samples. Diluents for complex matrices, such as serum, plasma, saliva, urine, and human milk, should be evaluated prior to use in this DuoSet.  
 

Product Features

• Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time

• Development protocols are provided to guide further assay optimization

• Assay can be customized to your specific needs

• Economical alternative to complete kits

Kit Content

• Capture Antibody

• Detection Antibody

• Recombinant Standard

• Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # ) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # ), or 0.05% Tween^® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # ), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # )

Stop Solution: 2 N H₂SO₄ (Catalog # )

Microplates: R&D Systems (Catalog # )

Plate Sealers: ELISA Plate Sealers (Catalog # )

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.

• Entrez Gene IDs:

  2638 (Human); 14473 (Mouse); 24384 (Rat);

• Long Name:

  Vitamin D Binding Protein

• Aliases:

  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