B7-2/CD86 Antibody (1029808) [Unconjugated]

Western blot shows lysates of Ramos human Burkitt's lymphoma cell line and human B7-2/CD86 knockout Ramos cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Cynomolgus Monkey B7‑2/CD86 Monoclonal Antibody (Catalog # MAB10601) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for B7‑2/CD86 at approximately 70 kDa (as indicated) in the parental cell line, but is not detectable in knockout cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Western blot shows lysates of HDLM‑2 human Hodgkin’s lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Cynomolgus Monkey B7‑2/CD86 Monoclonal Antibody (Catalog # MAB10601) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for B7‑2/CD86 at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

B7‑2/CD86 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Cynomolgus Monkey B7‑2/CD86 Monoclonal Antibody (Catalog # MAB10601) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in lymphocytes. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Pm-Cm
• Applications: WB, IHC, KO
• Clone: 1029808
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

B7-2/CD86 Antibody (1029808) [Unconjugated] Summary

• Immunogen: Human embryonic kidney cell HEK293-derived cynomolgus monkey B7-2/CD86 protein
  Leu20-His239
  Accession # XP_005548057
• Specificity: Detects cynomolgus monkey B7-2/CD86 in direct ELISAs.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 8-25 ug/mL
  • Knockout Validated 2 ug/mL
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for B7-2/CD86 Antibody (1029808) [Unconjugated]

• 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

Background

B7-2, also known as CD86, B70, and ETC-1, is a 60-100 kDa variably glycosylated protein in the B7 family. B7 family members are transmembrane cell surface molecules that play important roles in immune activation and the maintenance of immune tolerance (1, 2). Within the ECD, cynomolgus B7-2 shares 96%, 57%, and 56% amino acid sequence identity with human, mouse and rat B7-2, respectively. B7-2 is highly expressed on activated antigen presenting cells (APC), e.g. B cells, dendritic cells, and monocytes (2-5), as well as on vascular endothelial cells (6). B7-2 and the closely related B7-1/CD80 exhibit overlapping but distinct functional properties. Their binding to CD28, which is constitutively expressed on T cells, enhances T cell receptor signaling and also provides TCR-independent co-stimulation (3, 5, 7-9). B7-1 and B7-2 additionally bind the CD28-related protein, CTLA-4, which is up‑regulated and recruited to the immunological synapse (IS) at the onset of T cell activation (3, 5, 7, 8). CTLA-4 ligation inhibits the T cell response and supports regulatory T cell function (10). B7-2 is expressed earlier than B7-1 following APC activation (4), and both proteins bind with higher affinity to CTLA-4 than to CD28 (8). B7-2 promotes the stabilization of CD28 in the IS, while B7-1 is primarily responsible for promoting CTLA-4 recruitment and accumulation in the IS (11). The relative participation of B7-1 and B7-2 in T cell co-stimulation can also alter the Th1/Th2 bias of the immune response (12). Both B7-1 and B7-2 serve as cellular receptors for B species adenoviruses (13).

1. Greenwald, R.J. et al. (2005) Annu. Rev. Immunol. 23:515.
2. Bour-Jordan, H. et al. (2011) Immunol. Rev. 241:180.
3. Freeman, G.J. et al. (1993) J. Exp. Med. 178:2185.
4. Lenschow, D.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90:11054.
5. Hathcock, K.S. et al. (1993) Science 262:905.
6. Seino, K. et al. (1995) Int. Immunol.7:1331.
7. Chen, C. et al. (1994) J. Immunol. 152:4929.
8. Lanier, L.L. et al. (1995) J. Immunol. 154:97.
9. Rudd, C.E. et al. (2009) Immunol. Rev. 229:12.
10. Wing, K. et al. (2011) Trends Immunol. 32:428.
11. Pentcheva-Hoang, T. et al. (2004) Immunity 21:401.
12. Kuchroo, V.K. et al. (1995) Cell 80:707.
13. Short, J.J. et al. (2006) Virus Res. 122:144.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • XP_005548057()