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.
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Knockout Validated, Western Blot, Neutralization, Flow Cytometry, CyTOF-ready
Cited:
Neutralization, Flow Cytometry, Immunocytochemistry, ELISA Development, Functional Assay
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 37301
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Product Specifications
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human B7‑2/CD86
Ala23-His244
Accession # P42081
Ala23-His244
Accession # P42081
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.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human B7‑2/CD86 Antibody
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
Application
Recommended Usage
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Flow Cytometry
0.25 µg/106 cells
Sample: Human whole blood monocytes and Ramos lymphoma cell line
Sample: Human whole blood monocytes and Ramos lymphoma cell line
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
Western Blot
5 µg/mL
Sample: Daudi human Burkitt's lymphoma cell line
Sample: Daudi human Burkitt's lymphoma 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.
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
Protein A or G purified from ascites
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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.
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.
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
B72, CD86
Gene Symbol
CD86
UniProt
Additional B7-2/CD86 Products
Product Documents for Human B7‑2/CD86 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human B7‑2/CD86 Antibody
For research use only
Citations for Human B7‑2/CD86 Antibody
Customer Reviews for Human B7‑2/CD86 Antibody (1)
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