Thermo Fisher Scientific CD366 (TIM3) Monoclonal Antibody (RMT3-23), Brilliant Violet 711, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

0.5 µg/test

-

Product Specifications

Species Reactivity

Mouse

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), Brilliant Violet™ 711, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

RMT3-23

Conjugate

Brilliant Violet™ 711 Brilliant Violet™ 711 Brilliant Violet™ 711

View additional formats

• Unconjugated
• APC
• Brilliant Violet 421
• Brilliant Violet 605
• Brilliant Violet 650
• FITC
• PE
• PE-Cyanine7
• PerCP-eFluor 710
• Super Bright 436
• Super Bright 702
• Request custom conjugation

Excitation/Emission Max

406/711 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Wet ice

Product Specific Information

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™ 711 (BV711) is a tandem dye that emits at 713 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™ 711 (BV711) is not compatible with methanol-based fixation.

Excitation: 407 nm; Emission: 713 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.™

Target Information

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.