Measured by its ability to neutralize B7‑1/CD80-induced IL‑2 secretion in the Jurkat human acute T cell leukemia cell line. The Neutralization Dose (ND50) is typically 0.15-0.6 µg/mL in the presence of 0.1 µg/mL Recombinant Mouse B7‑1/CD80 Fc Chimera and 10 µg/mL PHA.
Please Note: Optimal dilutions should be determined by each laboratory for each application.
are available in the Technical Information section on our website.
Detection of Mouse B7‑1/CD80 by Western Blot.
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.
Detection of B7‑1/CD80 in Mouse Splenocytes by Flow Cytometry.
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.
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.
IL‑2 secretion Induced by B7‑1/CD80 and Neutralization by Mouse B7‑1/CD80 Antibody.
Recombinant Mouse B7‑1/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 Mouse B7‑1/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 typically 0.15‑0.6 µg/mL.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
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.
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Chen, C. et al. (1994) J. Immunol. 152:4929.
Freeman, G.J. et al. (1993) J. Exp. Med. 178:2185.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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