B7-1/CD80 Antibody [Unconjugated]

Recombinant MouseB7‑1/CD80 Fc Chimera (Catalog # 740-B1) co-stimulates IL‑2 secretion in the Jurkat human acute T cell leukemia cell line in the presence of PHA in a dose-dependent manner (orange line), as measured by the Human IL‑2 Quantikine ELISA Kit (Catalog # D2050). IL‑2 secretion elicited by Recombinant MouseB7‑1/CD80 Fc Chimera (0.1 µg/mL) and PHA (10 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740). The ND50 is typically0.15‑0.6 µg/mL.

B7‑1/CD80 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Mouse splenocytes either treated with 200 ng/mL LPS (filled histogram) or unstimulated (open histogram) were stained with Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Western blot shows lysates of C2C12 mouse myoblast cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse B7‑1/CD80 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF740) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for B7‑1/CD80 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

• Reactivity: Mu
• Applications: WB, Flow, B/N, CyTOF-ready, ELISA(Cap), ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

B7-1/CD80 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse B7‑1/CD80
  Asp37-Lys245
  Accession # Q00609
• Specificity: Detects mouse B7‑1/CD80 in ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CD80
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • CyTOF-ready
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • Immunocytochemistry 5-15 ug/mL
  • Neutralization 0.15-0.6 ug/mL
  • ELISA Standard (Matched Pair)
• Application Notes: ELISA Capture: Mouse B7-1/CD80 Antibody (Catalog # AF740)
  ELISA Detection: Mouse B7-1/CD80 Biotinylated Antibody (Catalog # BAF740)
  Standard: Recombinant Mouse B7-1/CD80 Fc Chimera (Catalog # 740-B1)

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 0.2 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 B7-1/CD80 Antibody [Unconjugated]

• Activation B7-1 antigen
• B7
• B71
• B7-1
• BB1
• CD28LG1
• CD28LGB7-1 antigen)
• CD80 antigen
• CD80 molecule
• CD80
• costimulatory factor CD80
• costimulatory molecule variant IgV-CD80
• CTLA-4 counter-receptor B7.1
• T-lymphocyte activation antigen CD80

Background

B7-1 and B7-2, together with their receptors CD28 and CTLA-4, constitute one of the dominant costimulatory pathways that regulate T- and B-cell responses. Although both CTLA-4 and CD28 can bind to the same ligands, CTLA-4 binds to B7-1 and B7-2 with a 20‑100 fold higher affinity than CD28 and is involved in the
down‑regulation of the immune response. B7-1 is expressed on activated B cells, activated T cells, and macrophages. B7-2 is constitutively expressed on interdigitating dendritic cells, Langerhans cells, peripheral blood dendritic cells, memory B cells, and germinal center B cells. Additionally, B7-2 is expressed at low levels on monocytes and can be up-regulated through interferon gamma. B7-1 and B7-2 are both members of the immunoglobulin superfamily. Mouse B7-1 is a 306 amino acid (aa) protein containing a putative 37 aa signal peptide, a 190 aa extracellular domain, a 22 aa transmembrane domain, and a 38 aa cytoplasmic domain. Mouse B7-1 and B7-2 share 28% amino acid identity. Mouse and human B7-1 share 44% amino acid identity. However, it has been observed that both human and mouse
B7‑1 and B7‑2 can bind to either human or mouse CD28 and CTLA-4, suggesting that there are conserved amino acids which form the B7-1/B7-2/CD28/CTLA-4 critical binding sites.

1. Azuma, M. et al. (1993) Nature 366:76.
2. Freeman, G.J. et al. (1993) Science 262:909.
3. Freeman, G. et al. (1991) J. Exp. Med. 174:625.
4. Selvakumar, A. et al. (1993) Immunogenetics 38:292.
5. Chen, C. et al. (1994) J. Immunol. 152:4929.
6. Freeman, G.J. et al. (1993) J. Exp. Med. 178:2185.

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 B7-1/CD80 Antibody (AF740)(4)

Publications using AF740

• VK Clements, T Long, R Long, C Figley, DMC Smith, S Ostrand-Ro High fat diet and leptin promote tumor progression by inducing myeloid-derived suppressor cells J. Leukoc. Biol., 2018;0(0):. 2018 [PMID: 29345342] (Flow Cytometry, Mouse)
• Garg S, Delaney C, Toubai T, Ghosh A, Reddy P, Banerjee R, Yung R Aging is associated with increased regulatory T-cell function. Aging Cell, 2014;13(3):441-8. 2014 [PMID: 24325345] (Western Blot, Mouse)
• Sofra V, Mansour S, Liu M, Gao B, Primpidou E, Wang P, Li S Antigen-loaded ER microsomes from APC induce potent immune responses against viral infection. Eur. J. Immunol., 2009;39(1):85-95. 2009 [PMID: 19089809] (Western Blot, Mouse)
• He H, Li W, Chen SY, Zhang S, Chen YT, Hayashida Y, Zhu YT, Tseng SC Suppression of activation and induction of apoptosis in RAW264.7 cells by amniotic membrane extract. Invest. Ophthalmol. Vis. Sci., 2008;49(10):4468-75. 2008 [PMID: 18614802] (ICC, Mouse)

Bioinformatics

• Gene Symbol: CD80
• Uniprot:
  • Q00609()