Detection of CTLA-4 in Mouse Splenocytes by Flow Cytometry.Mouse splenocytes treated with Concanavalin A for 48 hours were stained with Rat Anti-Mouse CD3 APC-conjugated Monoclonal Antibody (Catalog # FAB4841A) and either (A) Rat Anti-Mouse CTLA-4 Fluorescein-conjugated Monoclonal Antibody (Catalog # FAB434F) or (B) Rat IgG2A Fluorescein Isotype Control (Catalog # IC006F). View our protocol for Staining Membrane-associated Proteins.
CTLA-4 and CD28, together with their ligands B7-1 and B7-2, constitute one of the dominant costimulatory pathways that regulate T and B cell responses. CTLA-4 and CD28 are structurally homologous molecules that are members of the immunoglobulin (Ig) gene superfamily. Both CTLA-4 and CD28 are composed of a single Ig V‑like extracellular domain, a transmembrane domain and an intracellular domain. CTLA-4 and CD28 are both expressed on the cell surface as disulfide-linked homodimers or as monomers. The genes encoding these two molecules are closely linked on human chromosome 2. CTLA-4 was originally identified as a gene that was specifically expressed by cytotoxic T lymphocytes. However, CTLA-4 transcripts have since been found in both Th1 and Th2, and CD4+ and CD8+ T cell clones. Whereas, CD28 expression is constitutive on the surfaces of 95% of CD4+ T cells and 50% of CD8+ T cells and is down regulated upon T cell activation, CTLA-4 expression is upregulated rapidly following T cell activation and peaks approximately 24 hours following activation. Although both CTLA-4 and CD28 can bind to the same ligands, CTLA-4 binds to B7-1 and B7-2 with 20-100-fold higher affinity than CD28. The physiological role of CTLA-4 in T cell costimulation is currently being studied. Recombinant human or mouse CTLA-4/Fc chimera preparations produced at R&D Systems have been shown to bind both B7-1 and B7-2 with high affinity and to inhibit CD28 signalling competitively.
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