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Human B7-2/CD86 Antibody

Catalog #: MAB141
16 Citations 1 Review Datasheet / COA / SDS
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MAB141-SP
MAB141-100
MAB141-500

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Detection of Human B7‑2/CD86 by Western Blot.
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Citations (16)
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Human B7-2/CD86 Antibody Summary

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 either lyophilized or 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.

Scientific Data

Western Blot Detection of Human B7-2/CD86 antibody by Western Blot. View Larger

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.

Flow Cytometry Detection of B7-2/CD86 antibody in Human Blood Monocytes antibody by Flow Cytometry. View Larger

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 IgG1Isotype Control (Catalog # MAB002) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).

Flow Cytometry Detection of B7-2/CD86 antibody in Human Ramos Cell Line antibody by Flow Cytometry. View Larger

Detection of B7-2/CD86 in Human Ramos Cell Line by Flow Cytometry. Human Ramos lymphoma cell line was stained with Mouse Anti-Human B7 Formatting the figure legends (adjusted HTML for better line wrapping). Only applies 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.

Knockout Validated B7-2/CD86 Antibody Specificity is Shown by Flow Cytometry in Knockout Cell Line. View Larger

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.

Knockout Validated Western Blot Shows Human B7-2/CD86 Antibody Specificity by Using Knockout Cell Line. View Larger

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.

Neutralization Cell IL-2 Secretion Induced by B7‑2/CD86 and Neutralization by Human B7‑2/CD86 Antibody. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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

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Citations for Human B7-2/CD86 Antibody

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.

16 Citations: Showing 1 - 10
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  1. By Binding CD80 and CD86, the Vaccinia Virus M2 Protein Blocks Their Interactions with both CD28 and CTLA4 and Potentiates CD80 Binding to PD-L1
    Authors: Patricia Kleinpeter, Christelle Remy-Ziller, Eline Winter, Murielle Gantzer, Virginie Nourtier, Juliette Kempf et al.
    Journal of Virology
  2. Histamine promotes the development of monocyte-derived dendritic cells and reduces tumor growth by targeting the myeloid NADPH oxidase.
    Authors: Martner A, Wiktorin H, Lenox B, Ewald Sander F, Aydin E, Aurelius J, Thoren F, Stahlberg A, Hermodsson S, Hellstrand K
    J Immunol, 2015-04-13;194(10):5014-21.
  3. Immune atlas of pituitary neuroendocrine tumors highlights endocrine-driven immune signature and therapeutic implication
    Authors: Guo, X;Chang, M;Li, W;Qian, Z;Guo, H;Xie, C;Bi, WL;Xing, B;Zhang, F;Huang, Y;
    Cell reports
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. Ex vivo-expanded human CD19+TIM-1+ regulatory B cells suppress immune responses in vivo and are dependent upon the TIM-1/STAT3 axis
    Authors: S Shankar, J Stolp, SC Juvet, J Beckett, PS Macklin, F Issa, J Hester, KJ Wood
    Nature Communications, 2022-06-03;13(1):3121.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Analysis of transient membrane protein interactions by single-molecule diffusional mobility shift assay
    Authors: MG Jeong, K Zhou, S Park, H An, Y Kwon, Y Chang, DH Kim, SH Ryu
    Experimental & Molecular Medicine, 2021-02-19;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  6. Comparison of the biological characteristics of human mesenchymal stem cells derived from exfoliated deciduous teeth, bone marrow, gingival tissue, and umbilical cord
    Authors: J Li, SQ Xu, YM Zhao, S Yu, LH Ge, BH Xu
    Mol Med Rep, 2018-09-20;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  7. Endothelial extracellular vesicles modulate the macrophage phenotype: Potential implications in atherosclerosis
    Authors: S He, C Wu, J Xiao, D Li, Z Sun, M Li
    Scand. J. Immunol., 2018-04-01;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  8. Human trophoblast cells induced MDSCs from peripheral blood CD14(+) myelomonocytic cells via elevated levels of CCL2
    Authors: Xun Qu
    Cell. Mol. Immunol., 2015-06-01;13(5):615-27.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  9. 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, 2014-03-31;2014(0):593562.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  10. 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, 2012-09-12;72(22):5733-43.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  11. ICOS-LICOS interaction is critically involved in TGN1412-mediated T-cell activation.
    Blood, 2012-05-10;119(26):6268-77.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  12. 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, 2010-05-01;28(5):939-54.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  13. 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., 2010-03-01;40(3):770-9.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  14. 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., 2008-06-01;180(11):7287-93.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  15. 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., 2007-04-01;261(4):399-407.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Development
  16. 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, 2002-05-01;16(5):865-73.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Development

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Human B7‑2/CD86 Antibody
By Lu Bai on 10/11/2022
Application: Flow Sample Tested: Dendritic cells Species: Human