B7-H3/CD276 Antibody (185504) [Unconjugated]

Western blot shows lysates of U2OS human osteosarcoma parental cell line and B7-H3 knockout U2OS cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB1027) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for B7‑H3 at approximately 95 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human monocyte-derived dendritic cells were stained with Mouse Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB1027, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).

PC‑3 human prostate cancer cell line was stained with Mouse Anti-Human B7-H3 Monoclonal Antibody (Catalog # MAB1027, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B).

Western blot shows lysates of LNCaP human prostate cancer cell line and U2OS human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human B7‑H3 Monoclonal Antibody (Catalog # MAB1027) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for B7‑H3 at approximately 90-110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

• Reactivity: Hu
• Applications: WB, Flow, CyTOF-ready, KO
• Clone: 185504
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

B7-H3/CD276 Antibody (185504) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human B7‑H3
  Leu29-Pro245
  Accession # NP_079516
• Specificity: Detects human B7-H3 in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant human (rh) B7‑H1, rhB7‑H2, rhB7‑1, rhB7‑2, or recombinant mouse PD-L2 is observed.
• Details of Functionality: ActivityAssaywCitation not found None None None
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD276

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • CyTOF-ready
  • Knockout Validated
• Control:

  B7-H3/CD276 Knockout HeLa Cell Lysate

Packaging, Storage & Formulations

• 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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for B7-H3/CD276 Antibody (185504) [Unconjugated]

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

Background

Human B7 homolog 3 (B7-H3) is a member of the B7 family of immune proteins that provide signals for regulating immune responses (1‑3). Other family members include B7-1, B7-2, B7-H2, PD-L1 (B7-H1), and PD-L2. B7 proteins are immunoglobulin (Ig) superfamily members with extracellular Ig-V-like and Ig-C-like domains and short cytoplasmic domains. Among the family members, they share about 20‑40% amino acid (aa) sequence identity. The cloned human B7-H3 cDNA encodes a 316 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. An isoform of human B7-H3 containing a four-Ig-like domain extracellular region has also been identified. Human B7-H3 is not expressed on resting B cells, T cells, monocytes or dendritic cells, but is induced on dendritic cells and monocytes by inflammatory cytokines. B7-H3 expression is also detected on various normal tissues and in some tumor cell lines. Human B7-H3 does not bind any known members of the CD28 family of immunoreceptors. However, B7-H3 has been shown to bind an unidentified counter-receptor on activated T cells to costimulate the proliferation of CD4+ or CD8+ T cells. B7-H3 has also been found to enhance the induction of primary cytotoxic T lymphocytes and stimulate IFN-gamma production (1‑3).

Chapoval, A.I. et al. (2001) Nat. Immunol. 2:269.
Sharpe, A.H. and G.J. Freeman (2002) Nat. Rev. Immunol. 2:116.
Coyle, A. and J. Gutierrez-Ramos (2001) Nat. Immunol. 2:203.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for B7-H3/CD276 Antibody (MAB1027)(4)

Publications using MAB1027

• Sucker A, Zhao F, Real B, Heeke C, Bielefeld N, Mabetaen S, Horn S, Moll I, Maltaner R, Horn P, Schilling B, Sabbatino F, Lennerz V, Kloor M, Ferrone S, Schadendorf D, Falk C, Griewank K, Paschen A Genetic evolution of T-cell resistance in the course of melanoma progression. Clin Cancer Res, 2014;20(24):6593-604. 2014 [PMID: 25294904] (Flow Cytometry, Human)
• Ramsay, Alan G, Clear, Andrew J, Fatah, Rewas, Gribben, John G Multiple inhibitory ligands induce impaired T-cell immunologic synapse function in chronic lymphocytic leukemia that can be blocked with lenalidomide: establishing a reversible immune evasion mechanism in human cancer. Blood, 2012;120(7):1412-21. 2012 [PMID: 22547582] (Flow Cytometry, Human)
• Tran CN, Thacker SG, Louie DM, Oliver J, White PT, Endres JL, Urquhart AG, Chung KC, Fox DA Interactions of T cells with fibroblast-like synoviocytes: role of the B7 family costimulatory ligand B7-H3. J. Immunol., 2008;180(5):2989-98. 2008 [PMID: 18292521] (Flow Cytometry, Human)
• Das S, Suarez G, Beswick EJ, Sierra JC, Graham DY, Reyes VE Expression of B7-H1 on gastric epithelial cells: its potential role in regulating T cells during Helicobacter pylori infection. J. Immunol., 2006;176(5):3000-9. 2006 [PMID: 16493058] (Western Blot, Human)

Bioinformatics

• Gene Symbol: CD276
• Uniprot:
  • NP_079516()