Rat B7-2/CD86 Biotinylated Antibody Summary
Accession # NP_064466
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Rat B7-2/CD86 Sandwich Immunoassay
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.
Detection of B7‑2/CD86 in Rat Spleen. B7‑2/CD86 was detected in immersion fixed paraffin-embedded sections of Rat Spleen using Goat Anti-Rat B7‑2/CD86 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1340) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface and cytoplasm in splenocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Preparation and 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.
For optimal T cell expansion and activation, a signal induced by the engagement of the T cell receptor and a “co-stimulatory” signal(s) through distinct T cell surface molecules are required. Members of the B7 superfamily of counter-receptors were identified by their ability to interact with co-stimulatory molecules found on the surface of T cells. Members of the B7 superfamily are type I membrane proteins and include B7-1 (CD80), B7-2 (CD86), B7-H1 (PD-L1), B7-H2 (B7RP-1), B7-H3, and PD-L2 (1). B7-2 is expressed constitutively at low levels on most Antigen Presenting Cells (APC) and is rapidly upregulated upon cell activation (2). T cells express two different receptors (CD28 and CTLA-4) capable of binding both B7-1 and B7-2 (2). B7-2 binds to CD28 with the low affinity but binds to CTLA-4 with intermediate affinity. In contrast, B7-1 binds CD28 with intermediate affinity and CTLA-4 with high affinity. Additionally, these molecules have different kinetics for binding CD28 and CTLA-4 with B7-2 having a higher-binding dissociation kinetics (1). Engagement of CD28 by B7-2 increases T cell proliferation and IL-2, IL-4, and IFN-gamma production, thereby enhancing the immune response (3). In contrast, engagement of CTLA-4 is involved in the down-regulation of the immune response (4). Rat B7-2 cDNA encodes a 313 amino acid (aa) precursor protein containing a an extracellular domain, a transmembrane domain, and a cytoplasmic domain. Rat and human B7-1 share 54% aa identity.
- Coyle, A.J. and J-C. Gutierrez-Ramos (2001) Nature Immunol. 2:203.
- Sharpe, A.H. and G.J. Freeman (2002) Nature Reviews 2:116.
- Freeman, G.J. et al. (1995) Immunity 5:523.
- Walunas, T.L. et al. (1994) Immunity 1:405.
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