Thermo Fisher Scientific CD366 (TIM3) Monoclonal Antibody (8B.2C12), APC-eFluor 780, eBioscience

Applications

Tested Dilution

Publications

Western Blot (WB)

-

View 1 publication 1 publication

Flow Cytometry (Flow)

0.25 µg/test

View 1 publication 1 publication

Immunoprecipitation (IP)

-

View 1 publication 1 publication

Product Specifications

Published species

Not Applicable

Host/Isotype

Rat / IgG1, kappa

Recommended Isotype Control

Rat IgG1 kappa Isotype Control (eBRG1), APC-eFluor™ 780, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

8B.2C12

Conjugate

APC-eFluor™ 780 APC-eFluor™ 780 APC-eFluor™ 780

View additional formats

• Unconjugated
• Alexa Fluor 488
• APC
• Biotin
• eFluor 450
• Functional Grade
• PE
• PerCP-eFluor 710
• Super Bright 436
• Super Bright 600
• Super Bright 645
• Request custom conjugation

Excitation/Emission Max

756/785 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Wet ice

RRID

AB_2784710

Product Specific Information

Description: The 8B.2C12 monoclonal antibody reacts with mouse CD366 (TIM3), a Th1-specific cell surface protein. CD366 is a type I transmembrane protein and 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, but is absent on Th2 and Tc2. Other hematopoietic cell types, including naive T cells, B cells, macrophages and dendritic cells, do not express CD366, at least at the protein level. Expression of CD366 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. In this model, in vivo administration of 8B.2C12 resulted in a hyperacute and atypical disease phenotype. It is postulated that the engagement of CD366 during T cell activation results in the expansion and activation of macrophages and increased severity of autoimmune disease. The Tim gene family may have an important role in the regulation of autoimmunity and allergies.

The 8B.2C12 antibody binds to the BALB/c allele of CD366 while reactivity to the C57Bl/6 allele is significantly weaker.

Applications Reported: 8B.2C12 has been reported for use in flow cytometric analysis.

Applications Tested: This 8B.2C12 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.

APC-eFluor 780 emits at 780 nm and is excited with the Red laser (633-647 nm). 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 # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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: 633-647 nm; Emission: 780 nm; Laser: Red Laser.

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.