Description: The RMT3-23 monoclonal antibody reacts with mouse CD366 (TIM3), a Th1-specific cell surface protein. The RMT3-23 antibody reacts with CD366 protein expressed by both BALB/c and C57BL/6 strains of mice. CD366, a type I transmembrane protein, contains an immunoglobulin and a mucin-like domain in its extracellular portion and a tyrosine phosphorylation motif in its cytoplasmic portion. CD366 is expressed selectively by differentiated CD4^+Th1 and CD8^+Tc1 cells, but is absent on CD4^+Th2 and CD8^+Tc2 cells. Other hematopoietic cell types, including naive T cells, B cells, macrophages and dendritic cells, do not express CD366, at least at the protein level. CD366 expression is upregulated at a late stage of T cell differentiation on Th1 cells after 3 rounds of in vitro polarization suggesting a role for this molecule in the transport or effector function of Th1 cells rather than a contribution to T cell differentiation. In an experimental autoimmune encephalomyelitis (EAE) model, CD366 was shown to be expressed on most CD4^+ and CD8^+ T cells in the central nervous system at the onset of clinical signs of disease, while less than 2% of CD4^+ cells in the periphery expressed CD366 after immunization.
RMT3-23 has been shown to have functional activity; blocks DC recognition of apoptotic cells and also induces autoantibody production.
Applications Reported: This RMT3-23 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This RMT3-23 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
TIM3 (Hepatitis A virus cellular receptor 2, HAVCR2, T-cell immunoglobulin, mucin-dmain containing-3) is a 281 amino acid long, Type-1 Th1- specific cell surface glycoprotein expressed on terminally differentiated CD4+Th1 and CD8+Tc1 cells. TIM3 consists of an IgV-like domain, a mucin-like domain in the extracellular region, and a conserved Tyrosine phosphorylation motif in the cytoplasmic region. TIM3 is involved in macrophage activation and induction of autoimmune diseases. Further, TIM3 down-regulates aggressive Th1-mediated immune responses and facilitates in the development of immune tolerance. Pathological significance of TIM3 has been attributed to Experimental autoimmune encephalomyelitis (EAE), a Th-1 dependent autoimmune disease, and also enhances the severity of experimental autoimmune encephalomyelitis in mice.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD366; FLJ14428; HAVcr-2; Hepatitis A virus cellular receptor 2 homolog; sCD366; soluble CD366; soluble TIM 3; T-cell immunoglobulin and mucin domain containing 3; T-cell immunoglobulin and mucin domain-containing protein 3; T-cell immunoglobulin mucin receptor 3; T-cell membrane protein 3; TIM-3; TIMD-3
Gene Aliases: Havcr2; TIM-3; Tim3; Timd3
UniProt ID: (Mouse) Q8VIM0
Entrez Gene ID: (Mouse) 171285