Human B7-2/CD86 Antibody

(5 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human B7‑2/CD86 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross‑reactivity with recombinant mouse B7-2 and recombinant rat B7-2 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • 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
    0.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • 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. Linsley, P. et al. (1990) Proc. Natl. Acad. Sci. 87:5031. The Neutralization Dose (ND50) is typically 0.25-1.25 µg/mL in the presence of 2 µg/mL Recombinant Human B7‑2/CD86 Fc Chimera and 10 µg/mL PHA.
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 0.5 µg/mL of Goat Anti-Human B7‑2/CD86 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-141-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). 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.
Immunohistochemistry
B7‑2/CD86 in Human Tonsil.
B7‑2/CD86 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti‑Human B7‑2/CD86 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑141‑NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.
Immunohistochemistry
B7‑2/CD86 in Human Tonsil.
B7‑2/CD86 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human B7‑2/CD86 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-141-NA) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
IL‑2 secretion Induced by
B7‑2/CD86 and Neutralization by Human B7‑2/CD86 Antibody.
Recombinant Human B7‑2/CD86 Fc Chimera (Catalog # 141-B2) 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 Human B7‑2/CD86 Fc Chimera (2 µg/mL) and PHA (10 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human B7‑2/CD86 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-141-NA). The ND50 is typically
0.25‑1.25 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 0.5 µg/mL of Goat Anti-Human B7‑2/CD86 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-141-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). 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 0.2 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.

5 Citations: Showing 1 - 5
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Species
Applications
Sample Type
  1. Tumor microenvironment contributes to Epstein-Barr virus anti-nuclear antigen-1 antibody production in nasopharyngeal carcinoma
    Authors: P Ai, Z Li, Y Jiang, C Song, L Zhang, H Hu, T Wang
    Oncol Lett, 2017;14(2):2458-2462.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC paraffin embedded
  2. Regional immunity in melanoma: immunosuppressive changes precede nodal metastasis.
    Authors: Mansfield AS, Holtan SG, Grotz TE
    Mod. Pathol., 2011;24(4):487-94.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. Modulation of T-cell activation by malignant melanoma initiating cells.
    Authors: Schatton T, Schutte U, Frank NY, Zhan Q, Hoerning A, Robles SC, Zhou J, Hodi FS, Spagnoli GC, Murphy GF, Frank MH
    Cancer Res., 2010;70(2):697-708.
    Species: Human
    Sample Type: Whole Tissue
    Application: IF
  4. 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
  5. 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
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