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Key Product Details

Validated by

Independent Antibodies

Species Reactivity

Validated:

Human

Cited:

Human hepegivirus

Applications

Validated:

CyTOF-ready, Flow Cytometry, Knockout Validated, Neutralization, Western Blot

Cited:

ELISA Development, Flow Cytometry, Functional Assay, Immunocytochemistry, Neutralization

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 37301

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human B7‑2/CD86
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 antibody by Western Blot.

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 antibody in Human Blood Monocytes antibody by Flow Cytometry.

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 antibody in Human Ramos Cell Line antibody by Flow Cytometry.

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.

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

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

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.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Reconstitution Buffer Available:
Size / Price
Qty
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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

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.

Alternate Names

B72, CD86

Entrez Gene IDs

942 (Human); 12524 (Mouse); 56822 (Rat); 102147235 (Cynomolgus Monkey)

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 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

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