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.
Human B7-2/CD86 Antibody (37301)
R&D Systems | Catalog # MAB141
Scientific images may be resized, cropped, or adjusted for brightness or contrast for presentation purposes. These updates are limited to presentation and do not affect the underlying scientific interpretation of the data. Where available, additional source data may be provided upon request.
Key Product Details
Validated by
Species Reactivity
Validated:
Cited:
Applications
Validated:
Cited:
Label
Antibody Source
Product Specifications
Immunogen
Ala23-His244
Accession # P42081
Specificity
Clonality
Host
Isotype
Endotoxin Level
Scientific Data Images for Human B7-2/CD86 Antibody (37301)
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 IgG1Isotype 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 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.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.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.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.Applications for Human B7-2/CD86 Antibody (37301)
CyTOF-ready
Flow Cytometry
Sample: Human whole blood monocytes and Ramos lymphoma cell line
Knockout Validated
Western Blot
Sample: Daudi human Burkitt's lymphoma cell line
Neutralization
Reviewed Applications
Read 1 review rated 4 using MAB141 in the following applications:
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & Storage
- 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.
Calculators
Background: B7-2/CD86
References
- 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.
Alternate Names
Gene Symbol
UniProt
Additional B7-2/CD86 Products
Product Documents for Human B7-2/CD86 Antibody (37301)
Certificate of Analysis
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Product Specific Notices for Human B7-2/CD86 Antibody (37301)
For research use only
Citations for Human B7-2/CD86 Antibody (37301)
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Application: Flow CytometrySample Tested: Dendritic cellsSpecies: HumanVerified Customer | Posted 10/11/2022
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- Protocol for Liperfluo
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- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars