Detects human B7‑2/CD86 in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human (rh) B7-1, recombinant mouse B7-2, recombinant rat B7-2, rhB7-H1, rhB7-H2, rhB7-H3, rhB7-H3b, rhB7-H4, or rhB7-L2 is observed.
Monoclonal Mouse IgG1 Clone # 37301
Protein A or G purified from ascites
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human B7‑2/CD86 Ala23-His244 Accession # P42081
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
B7-2/CD86 is specifically detected in Ramos human Burkitt's lymphoma parental cell line but is not detectable in B7-2/CD86 knockout Ramos cell line
Measured by its ability to neutralize B7‑2/CD86-induced IL-2 secretion in the Jurkat human acute T cell leukemia cell line. Freeman, G.J. et al. (1993) Science 262:909. The Neutralization Dose (ND50) is typically 0.5-2.5 μg/mL in the presence of 2 μg/mL Recombinant Human B7‑2/CD86 Fc Chimera.
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 Human B7‑2/CD86 by Western Blot.
Western blot shows lysates of Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 5 µg/mL of Mouse Anti-Human B7‑2/CD86 Monoclonal Antibody (Catalog # MAB141) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for B7‑2/CD86 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of B7‑2/CD86 in Human Blood Monocytes by Flow Cytometry.
Human peripheral blood monocytes were stained with Mouse Anti-Human CD14 PE‑conjugated Monoclonal Antibody (Catalog # FAB3832P) and either (A) Mouse Anti-Human B7‑2/CD86 Monoclonal Antibody (Catalog # MAB141) or (B) Mouse IgG1 Isotype Control (Catalog # MAB002) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).
Detection of B7-2/CD86 in Human Ramos Cell Line by Flow Cytometry.
Human Ramos lymphoma cell line was stained with Mouse Anti-Human B7Formatting the figure legends (adjusted HTML for better linewrapping). Onlyapplies to the website insert.2/CD86 Monoclonal Antibody (Catalog # MAB141, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by PE-conjugated Goat Anti-Mouse IgG secondary antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.
Western Blot Shows Human B7-2/CD86 Specificity by Using Knockout Cell Line.
Western blot shows lysates of Ramos human Burkitt's lymphoma parental cell line and B7-2/CD86 knockout Ramos cell line (KO). PVDF membrane was probed with 5 µg/mL of Mouse Anti-Human B7‑2/CD86 Monoclonal Antibody (Catalog # MAB141) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for B7‑2/CD86 at approximately 74 kDa (as indicated) in the parental Ramos cell line, but is not detectable in knockout Ramos cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
B7-2/CD86 Specificity is Shown by Flow Cytometry in Knockout Cell Line.
B7‑2/CD86 knockout Ramos human lymphoma cell line was stained with Mouse Anti-Human B7‑2/CD86 Monoclonal Antibody (Catalog # MAB141, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by PE-conjugated Goat anti-Mouse IgG Secondary Antibody (Catalog # F0102B). No staining in the B7‑2/CD86 knockout Ramos cell line was observed. View our protocol for Staining Membrane-associated Proteins.
Cell IL-2 Secretion Induced by B7‑2/CD86 and Neutralization by Human B7‑2/CD86 Antibody.
Recombinant Human B7‑2/CD86 Fc Chimera induces IL‑2 secretion in the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line), as measured by the Human IL-2 Quantikine kit (Catalog # D2050). Under these conditions, IL-2 secretion elicited by B7‑2/ CD86 is neutralized (green line) by increasing concentrations of Mouse Anti-Human B7‑2/CD86 Monoclonal Antibody (Catalog # MAB141). The ND50 is typically 0.5-2.5 μg/mL.
Preparation and Storage
Reconstitute at 0.5 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. Human B7-2 is a 329 amino acid (aa) protein containing a putative 23 aa signal peptide, a 224 aa extracellular domain, a 21 aa transmembrane domain, and a 61 aa cytoplasmic domain. Human B7-2 and B7-1 share 26% amino acid identity. Human and mouse B7-2 share 50% 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.
Azuma, M. et al. (1993) Nature 366:76.
Freeman, G.J. et al. (1993) Science 262:909.
Freeman, G. et al. (1991) J. Exp. Med. 174:625.
Selvakumar, A. et al. (1993) Immunogenetics 38:292.
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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