Human B7-2/CD86 Antibody

(8 citations)   
  • Species Reactivity
    Human
  • Specificity
    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.
  • Source
    Monoclonal Mouse IgG1 Clone # 37301
  • Purification
    Protein A or G purified from ascites
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human B7‑2/CD86
    Ala23-His244
    Accession # P42081
  • Formulation
    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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    5 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Knockout Validated
    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
  • Neutralization
    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. General Protocols are available in the Technical Information section on our website.
Data Examples
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).
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.

Knockout Validated
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    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.
Background: B7-2/CD86

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.

  • References:
    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.
  • Entrez Gene IDs:
    942 (Human); 12524 (Mouse); 56822 (Rat); 102147235 (Cynomolgus Monkey)
  • Alternate Names:
    Activation B7-2 antigen; B70; B7-2 antigen; B72; B7-2; B-lymphocyte activation antigen B7-2; BU63; CD28 antigen ligand 2; CD28LG2B7-2 antigen); CD86 antigen; CD86 molecule; CD86; CTLA-4 counter-receptor B7.2; FUN-1; LAB72; MGC34413; T-lymphocyte activation antigen CD86
Related Research Areas
Citations:

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.

8 Citations: Showing 1 - 8
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Species
Applications
Sample Type
  1. Tumor-activated TCRgammadelta(+) T cells from gastric cancer patients induce the antitumor immune response of TCRalphabeta(+) T cells via their antigen-presenting cell-like effects.
    Authors: Mao C, Mou X, Zhou Y, Yuan G, Xu C, Liu H, Zheng T, Tong J, Wang S, Chen D
    J Immunol Res, 2014;2014(0):593562.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  2. Endoneurial macrophages induce perineural invasion of pancreatic cancer cells by secretion of GDNF and activation of RET tyrosine kinase receptor.
    Authors: Cavel O, Shomron O, Shabtay A, Vital J, Trejo-Leider L, Weizman N, Krelin Y, Fong Y, Wong R, Amit M, Gil Z
    Cancer Res, 2012;72(22):5733-43.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fresh
  3. ICOS-LICOS interaction is critically involved in TGN1412-mediated T-cell activation.
    Blood, 2012;119(26):6268-77.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  4. Critical role of lipid rafts in CD154-mediated T cell signaling.
    Authors: El Fakhry Y, Alturaihi H, Diallo D, Merhi Y, Mourad W
    Eur. J. Immunol., 2010;40(3):770-9.
    Species: Human
    Sample Type: Whole Cells
    Application: Flow
  5. Phagocytosis of apoptotic cells modulates mesenchymal stem cells osteogenic differentiation to enhance IL-17 and RANKL expression on CD4+ T cells.
    Authors: Tso GH, Law HK, Tu W
    Stem Cells, 2010;28(5):939-54.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  6. Circulating myeloid dendritic cells of advanced cancer patients result in reduced activation and a biased cytokine profile in invariant NKT cells.
    Authors: van der Vliet HJ, Wang R, Yue SC, Koon HB, Balk SP, Exley MA
    J. Immunol., 2008;180(11):7287-93.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  7. The soluble forms of CD28, CD86 and CTLA-4 constitute possible immunological markers in patients with abdominal aortic aneurysm.
    Authors: Sakthivel P, Shively V, Kakoulidou M
    J. Intern. Med., 2007;261(4):399-407.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
  8. Human plasma contains a soluble form of CD86 which is present at elevated levels in some leukaemia patients.
    Authors: Hock BD, Patton WN, Budhia S, Mannari D, Roberts P, McKenzie JL
    Leukemia, 2002;16(5):865-73.
    Species: Human
    Sample Type: Plasma
    Application: ELISA Development
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
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Mouse IgG1 Isotype Control

Ctrl MAB002R 1
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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