Detection of TIM‑3 in Th1-stimulated Human PBMCs by Flow Cytometry.
Unstimulated (A) and Th1-stimulated (B) human peripheral blood mononuclear cells (PBMCs) were stained with Rat Anti-Human TIM‑3 Alexa Fluor® 594‑conjugated Monoclonal Antibody (Catalog # FAB2365T) and Mouse Anti-Human CD4 APC‑conjugated Monoclonal Antibody (Catalog # FAB3791A). Quadrant markers were set based on control antibody staining (Catalog # IC006T). 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.
TIM-3 (T cell Immunoglobulin and Mucin domain-3), also known as HAVCR2, is a 60 kDa member of the TIM family of immune regulating molecules. TIMs are type I transmembrane glycoproteins with one Ig-like V-type domain and a Ser/Thr-rich mucin stalk region (1, 2). Mature human TIM-3 consists of a 181 amino acid (aa) extracellular domain (ECD), a 21 aa transmembrane segment, and a 78 aa cytoplasmic tail (3). An alternatively spliced isoform is truncated within the mucin-like stalk. Within the ECD, human TIM-3 shares 58% aa sequence identity with mouse and rat TIM-3. TIM-3 is up‑regulated on several populations of activated myeloid cells (macrophages, monocytes, dendritic cells, microglia, and mast cells) and T cells (Th1, CD8+, NK, and Treg) (3-10). Its binding to Galectin-9 induces a range of immunosuppressive functions which enhance immune tolerance and inhibit anti-tumor immunity (11). TIM-3 ligation attenuates CD8+ and Th1 cell responses (11-13) and promotes the activity of Treg and myeloid derived suppressor cells (8, 11, 13, 14). In addition, dendritic cell-expressed TIM-3 dampens inflammation by enabling the phagocytosis of apoptotic cells and the cross-presentation of apoptotic cell antigens (4). It also binds the alarmin HMGB1, thereby preventing the activation of TLRs in response to released tumor cell DNA (7). TIM-3 interactions with Galectin-9 can alternatively trigger immune stimulatory effects, such as the coactivation of NK cell cytotoxicity (10).
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