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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 0.5 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.
Brilliant Violet™ 605 (BV605) is a tandem dye that emits at 605 nm and is intended for use on cytometers equipped with a violet (405 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright, Brilliant Violet™, Brilliant Ultra Violet™, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.
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 # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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.
Our internal testing suggests that Brilliant Violet™ 605 (BV605) is not compatible with methanol-based fixation.
Excitation: 407 nm; Emission: 605 nm; Laser: Violet Laser.
BRILLIANT VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen.™
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.
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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
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