• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Range

  0.78 - 50 ng/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.

Ancillary Reagent Kit Available  

DY008, DuoSet ELISA Ancillary Reagent Kit 2 -

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant humanB7 Homolog 3 (B7-H3). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, and urine samples. Diluents for complex matrices, such as serum, plasma, and urine, 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: B7-H3

Human B7-H3 was originally identified as a 316 amino acid (aa) type I membrane precursor protein with a putative 28 aa signal peptide, a 217 aa extracellular region containing one V-like and one C-like Ig domain, a transmembrane region, and a 45 aa cytoplasmic domain. Subsequently, a second dominantly expressed form of human B7-H3, containing a splice variation that duplicates the V-like and C-like Ig domains, was found. The 534 aa splice variant of B7-H3 has been referred to as B7-H3b, 4Ig-B7-H3, and B7-H3VCVC. RTPCR transcripts for both B7-H3 and 4Ig-B7-H3 have been found in all tissues except peripheral blood leukocytes. Southern blot analysis indicates that the 4Ig-B7-H3 isoform of B7-H3 is the predominate isoform expressed in human tissues.

In mouse, only a single form of B7-H3 containing one V-like and one C-like Ig domain was detected. Mouse and human B7-H3 share 87% aa sequence identity. B7-H3 has been shown to be expressed at very high levels in immature dendritic cells, at moderate levels on mature dendritic cells, LPS stimulated immature dendritic cells and LPS stimulated monocytes, and at low levels on resting monocytes. B7-H3 binds to activated T cells via an as yet unidentified receptor.

• Entrez Gene IDs:

  80381 (Human); 102657 (Mouse); 102131295 (Cynomolgus Monkey);

• Long Name:

  B7 Homolog 3

• Aliases:

  B7H3; B7-H3; B7H34Ig-B7-H3; B7-H3B7 homolog 3; CD276 antigen; CD276 molecule; CD276; Costimulatory molecule