Recombinant Human B7-H3 Fc Chimera Avi-tag Protein, CF

R&D Systems, part of Bio-Techne | Catalog # AVI1027

Biotinylated
R&D Systems, part of Bio-Techne
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AVI1027-050
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Key Product Details

Learn more about Avi-tag Biotinylated Proteins

Source

CHO

Accession #

NP_079516.1

Structure / Form

Disulfide-linked homodimer, biotinylated via Avi-tag

Conjugate

Biotin

Applications

Bioactivity

View all B7-H3 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human B7-H3 protein
Human B7-H3
(Leu29-Pro245)
Accession # NP_079516.1		 DIEGRMD Human IgG1
(Pro100-Lys330)		 Avi-tag
N-terminus C-terminus

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu29

Predicted Molecular Mass

52 kDa

SDS-PAGE

62-74 kDa, under reducing conditions

Activity

Measured by its ability to inhibit anti-CD3 antibody induced IL-2 or IFN-gamma secretion by human T cells.
The ED50 for this effect is 1-10 µg/mL.

Measured by its binding ability in a functional ELISA.
When Human B7-H3 Antibody (Catalog # AF1027) is immobilized at 1 μg/mL, 100 μL/well, the concentration of Biotinylated Recombinant Human B7-H3 Fc Chimera Avi-tag (Catalog # AVI1027) that produces 50% of the optimal binding response is approximately 3-18 ng/mL.

Scientific Data Images for Recombinant Human B7-H3 Fc Chimera Avi-tag Protein, CF

Recombinant Human B7-H3 Fc Chimera Avi-tag Protein Binding Activity.

When Human B7-H3 Antibody (AF1027) is immobilized at 1 μg/mL, 100 μL/well, the concentration of Biotinylated Recombinant Human B7-H3 Fc Chimera Avi-tag (Catalog # AVI1027) that produces 50% of the optimal binding response is approximately 3-18 ng/mL.

Biotinylated Recombinant Human B7-H3 Fc Chimera Avi-tag SDS-PAGE.

2 μg/lane of Biotinylated Recombinant Human B7-H3 Fc Chimera Avi-tag Protein (Catalog # AVI1027) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 62-74 kDa and 120-150 kDa, respectively.

Formulation, Preparation and Storage

AVI1027
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: B7-H3

T cells require a signal induced by the engagement of the T cell receptor and a "co-stimulatory" signal(s) through distinct T cell surface molecules for optimal T cell expansion and activation. Members of the B7 superfamily of counter-receptors were identified by their ability to interact with co-stimulatory molecules found on the surface of T cells. Members of the B7 superfamily include B7-1 (CD80), B7-2 (CD86), B7-H1 (PD-L1), B7-H2 (B7RP-1), PD-L2 (B7-DC), and B7-H3 (1). 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 (2). 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 (3𔂯). RT-PCR transcripts for both B7-H3 and 4Ig‑B7‑H3 have been found in all tissues except peripheral blood leukocytes (5). Southern blot analysis indicates that the 4Ig-B7-H3 isoform of B7‑H3 is the predominate isoform expressed in human tissues (5). In mouse, only a single form of B7-H3 containing one V-like and one C-like Ig domain was detected (5). Mouse and human B7-H3 share 87% aa sequence identity (3). 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 (3). B7-H3 binds to activated T cells via an as yet unidentified receptor. In assays using sub-optimal amount so anti-CD3 stimulation, 2Ig‑B7‑H3 enhances T cell proliferation, T cell interferon-gamma (IFN‑gamma ) production, and cytotoxic T cells induction (2). In an assay in which cells were transfected with 4Ig-B7-H3 or 2Ig‑B7‑H3 and anti-CD3 antibody was added, neither 4Ig-B7-H3 nor 2Ig‑B7‑H3 was capable of co‑stimulating T cell proliferation or IFN-gamma production (4, 5). Our Avi-tag Biotinylated human B7-H3 features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

References

  1. Coyle, A.J. and J.-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203.
  2. Chapoval, A.I. et al. (2001) Nature Immunol. 2:269.
  3. Sun, M. et al. (2002) J. Immunol. 168:6294.
  4. Steinberger, et al. (2004) J. Immunol. 172:2352.
  5. Ling, V. et al. (2003) Genomic 82:365.

Long Name

B7 Homolog 3

Alternate Names

B7H3, CD276

Entrez Gene IDs

80381 (Human); 102657 (Mouse); 315716 (Rat); 102131295 (Cynomolgus Monkey)

Gene Symbol

CD276

UniProt

NP_079516.1

Additional B7-H3 Products

Product Documents for Recombinant Human B7-H3 Fc Chimera Avi-tag Protein, CF

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Product Specific Notices for Recombinant Human B7-H3 Fc Chimera Avi-tag Protein, CF

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