Detection of CTLA‑4 in Mouse Splenocytes by Flow Cytometry.
Mouse splenocytes either untreated (A) or treated (B) with Conconavalin A for 72 hours were stained with Rat Anti-Mouse CTLA‑4 APC‑conjugated Monoclonal Antibody (Catalog # FAB434A) and Rat Anti-Mouse CD3 PE‑conjugated Monoclonal Antibody (Catalog # FAB4841P). Quadrant markers were set based on control antibody staining (Catalog # IC006A). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
CTLA-4 (also known as CD152) 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 is expressed on the cell surface as either a disulfide-linked homodimer or a 24-26 kDa monomer. 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 contextually defined. Full-length CTLA-4, when induced and activated, sends an inhibitory signal to the expressing T cell. It also promotes the development of Tc17 cells. Tregs constitutively express CTLA-4, a phenotype that is essential to their function. Over amino acids (aa) 36-161, mouse CTLA-4 shares 94% and 67% aa sequence identity with rat and human CTLA-4, respectively.
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