CD366 (TIM3) Monoclonal Antibody (RMT3-23), PerCP-eFluor™ 710, eBioscience™, Invitrogen™

Rat Monoclonal Antibody

Supplier:  LIFE TECHNOLOGIES 46587082

Catalog No. 46-587-082

Description

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 may 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. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. 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 (Product No. 00-8222-49) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333-57) 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 nm; Emission: 710 nm; Laser: Blue Laser 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.

Specifications

• Antigen: CD366 (TIM3)
• Classification: Monoclonal
• Concentration: 0.2 mg/mL
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene Accession No.: Q8VIM0
• Gene Symbols: Havcr2
• Purification Method: Affinity Chromatography
• Regulatory Status: RUO
• Gene ID (Entrez): 171285
• Content And Storage: 4° C, store in dark, DO NOT FREEZE!
• Form: Liquid

• Applications: Flow Cytometry
• Clone: RMT3-23
• Conjugate: PerCP-eFluor 710
• Gene: Havcr2
• Gene Alias: Havcr2; TIM-3; Tim3; Timd3
• Host Species: Rat
• Quantity: 100 μg
• Primary or Secondary: Primary
• Target Species: Mouse
• Product Type: Antibody
• Isotype: IgG2a κ