Detection of TIGIT in Mouse splenocytes by Flow Cytometry.
Mouse splenocytes treated with 50 ng/mL PMA and 200 ng/mL Calcium Ionomycin overnight were stained with Rat Anti-Mouse CD4 APC‑conjugated Monoclonal Antibody (Catalog # FAB554A) and either (A) Sheep Anti-Mouse TIGIT Alexa Fluor® 488‑conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB7267G) or (B) Normal Sheep IgG Allophycocyanin Control (Catalog # IC016A). 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.
TIGIT (T cell Immunoreceptor with Ig and ITIM domains; also Vstm3 and Vsig9) is a 30-34 kDa member of the CD28 family, Ig superfamily of molecules. It is expressed by NK cells and multiple subsets of mature T cells, and binds to either PVR/CD155 or PVRL2/CD112 that appear on dendritic cells (DC) and endothelium. Along with CD226, TIGIT and CD226/DNAM1 pairings with PVR and PVRL2 appear to form a network that resembles the well-characterized B7-1 and B7-2 pairings with CD28 and CTLA4 system. Binding of TIGIT by DC induces DC IL-10 release and inhibits IL-12 production. Ligation of TIGIT on T cells dampens TCR-mediated activation, while NK cell TIGIT ligation blocks NK cell cytotoxicity and IFN-gamma production. Mature mouse TIGIT is a type I transmembrane protein 215 amino acids (aa) in length. It contains a 116 aa extracellular region (aa 26-141) with a V-type Ig-like domain (aa 27-125), and a 79 aa cytoplasmic domain with one ITIM motif. There is one isoform variant that is unusual in that it shows an addition of nine amino acids spread over three insertion sites. Over aa 26-143, mouse TIGIT shares only 84% aa sequence identity with rat TIGIT, but only 68% aa sequence identity with human TIGIT.
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