B7-H3/CD276 Antibody [Unconjugated]

Western blot shows lysates of MEF mouse embryonic feeder cells, P19 mouse embryonal carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, C2C12 mouse myoblast cell line, and 3T3-L1 mouse embryonic fibroblast adipose-like cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse B7-H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1397) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7-H3 at approximately 45-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Recombinant Mouse B7‑H3 enhances proliferation in mouse CD3+ T cells in the presence of 100 ng/mL Hamster Anti-Mouse CD3 epsilon Monoclonal Antibody (Catalog # MAB484) in a dose-dependent manner (orange line), as measured by the Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Mouse B7‑H3 (2 μg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse B7‑H3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1397). The ND50 is typically 1.5-7.5 μg/mL.

• Reactivity: Mu
• Applications: WB, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

B7-H3/CD276 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse B7‑H3
  Val29-Phe244
  Accession # Q8VE98
• Specificity: Detects mouse B7‑H3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 50% cross‑reactivity with recombinant human (rh) B7-H3 is observed, and less than 1% cross-reactivity with recombinant mouse (rm) B7‑H1, rmB7-H2 and rmB7-1 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CD276
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Neutralization 0.15-0.6 ug/mL
  • Western Blot 1 ug/mL

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 either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 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 [Unconjugated]

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

Background

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), B7-H3, and PD-L2 (1). B7-H3 is 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-of-yet identified receptor. B7-H3 co-stimulates proliferation of T cells and interferon-gamma (IFN-gamma ) production and enhances the induction of cytotoxic T cells. B7-H3 shares 20‑27% amino acid (aa) identity with other B7 family members (2). Murine B7-H3 is a 259 aa protein containing an extracellular domain, a transmembrane domain and a cytoplasmic domain. Mouse and human B7-H3 share 87% aa identity (3).

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.

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 (AF1397)(4)

Publications using AF1397

• Liu HJ, Du H, Khabibullin D et al. mTORC1 upregulates B7-H3/CD276 to inhibit antitumor T cells and drive tumor immune evasion Nature communications 3/3/2023 [PMID: 36869048]
• Liu HJ, Du H, Khabibullin D et al. mTORC1 upregulates B7-H3/CD276 to inhibit antitumor T cells and drive tumor immune evasion Nature communications 2023-03-03 [PMID: 36869048]
• E Picarda, PM Galbo, H Zong, MR Rajan, V Wallenius, D Zheng, E Börgeson, R Singh, J Pessin, X Zang The immune checkpoint B7-H3 (CD276) regulates adipocyte progenitor metabolism and obesity development Science Advances, 2022-04-27;8(17):eabm7012. 2022-04-27 [PMID: 35476450] (Western Blot, Mouse)
• Sun J, Fu F, Gu W, Yan R, Zhang G, Shen Z, Zhou Y, Wang H, Shen B, Zhang X Origination of new immunological functions in the costimulatory molecule B7-H3: the role of exon duplication in evolution of the immune system. PLoS ONE, 2011-09-13;6(9):e24751. 2011-09-13 [PMID: 21931843] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: CD276
• Uniprot:
  • Q8VE98()