APC Rabbit anti-Mouse CD366 (TIM-3) Monoclonal Antibody for Flow Cytometry - Q8VIM0

Core Product Specifications and Parameters

Biological Background: CD366 (TIM-3) Function and Localization

• Hepatitis A virus cellular receptor 2 homolog (HAVCR2), also known as T-cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) or T-cell membrane protein 3, is a type I transmembrane glycoprotein belonging to the immunoglobulin superfamily. Alternative splicing produces isoforms, and the canonical mouse sequence has a calculated molecular weight of approximately 31 kDa (22 kDa observed due to glycosylation).
• Functions as an immunomodulatory receptor with generally inhibitory roles, suppressing T-helper type 1 (Th1)-mediated auto- and alloimmune responses and promoting immunological tolerance (PubMed:14556006, PubMed:18006747). In CD8+ T cells, it attenuates TCR signaling by blocking NF-kappaB and NFAT promoter activities, reducing IL-2 secretion (By similarity). Related references: PMID:14556006 PMID:18006747
• However, context-dependent activating functions have been reported, including enhancement of TCR signaling in T cells via ZAP70, LCP2, LCK, and FYN (PubMed:21807895) and boosting CD8+ T-cell responses during acute infection (PubMed:24567532). On natural killer (NK) cells, it acts as a coreceptor to enhance IFN-gamma production but can also suppress cytotoxicity depending on conditions (PubMed:25337993). Related references: PMID:21807895 PMID:24567532 PMID:25337993
• TIM-3 is a receptor for LGALS9 (galectin-9). This interaction leads to suppression of T-cell responses and apoptosis through HAVCR2 phosphorylation and dissociation from BAG6 (PubMed:22863785). It also contributes to innate immunity by stimulating macrophage IL-1β secretion and restricting intracellular bacterial growth upon binding to LGALS9 on Mycobacterium tuberculosis-infected macrophages (PubMed:20937702). Related references: PMID:22863785 PMID:20937702
• Additionally, TIM-3 binds phosphatidylserine (PtSer) in a calcium-dependent manner, mediating phagocytosis of apoptotic cells by dendritic cells (PubMed:20083673, PubMed:19224762). In tumor-infiltrating dendritic cells, it suppresses nucleic acid-mediated innate immune responses by interacting with HMGB1 (PubMed:22842346). Related references: PMID:20083673 PMID:19224762 PMID:22842346
• Subcellular localization includes the plasma membrane, cell junctions, and a secreted form. Post-translational modifications include disulfide bonds, glycosylation, and phosphorylation, which regulate its function and signaling interactions.

Experimental Guidance and Technical Tips

• The immunogen corresponds to the extracellular domain (amino acids 20-191), ensuring recognition of the native receptor on the cell surface, suitable for flow cytometry.
• Consider titrating the antibody to determine optimal staining concentration for your specific cell type, as TIM-3 expression levels vary with activation status.
• Since TIM-3 is expressed on activated T cells, macrophages, dendritic cells, NK cells, and mast cells, validate the antibody in the relevant sample system and include appropriate isotype controls.
• For flow cytometry, use a viability dye to exclude dead cells, as TIM-3 can bind phosphatidylserine on apoptotic cells, potentially causing non-specific staining.

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