Thermo Fisher Scientific CD366 (TIM3) Monoclonal Antibody (8B.2C12), Super Bright 645, eBioscience

Applications and Tested Dilution

Product Specifications

Additional Available Formats

• Unconjugated
• Alexa Fluor 488
• APC
• APC-eFluor 780
• Biotin
• eFluor 450
• Functional Grade
• PE
• PerCP-eFluor 710
• Super Bright 436
• Super Bright 600
• Request custom conjugation

Product Specific Information

The 8B.2C12 monoclonal antibody reacts with mouse CD366 (TIM3), a Th1-specific cell surface protein. CD366 is a type I transmembrane protein containing an immunoglobulin and mucin-like domain extracellularly, and a tyrosine phosphorylation motif intracellularly. It is selectively expressed on differentiated CD4⁺ Th1 and CD8⁺ Tc1 cells, but absent on Th2 and Tc2 cells. Other hematopoietic cells such as naive T cells, B cells, macrophages, and dendritic cells do not express CD366 at the protein level.

Expression of CD366 increases during late Th1 differentiation, indicating a role in effector function rather than differentiation. In EAE models, CD366 expression was observed on most CD4⁺ and CD8⁺ T cells in the CNS at disease onset, while peripheral expression remained low. In vivo administration of 8B.2C12 induced a hyperacute and atypical disease phenotype, suggesting CD366 engagement enhances macrophage activation and autoimmune severity.

The 8B.2C12 antibody binds strongly to the BALB/c allele of CD366, with weaker reactivity to the C57Bl/6 allele.

Applications Reported: Flow cytometric analysis
Applications Tested: ≤0.125 µg/test (100 µL final volume; 10⁵–10⁸ cells/test)
Optimize antibody concentration empirically for best results.

Fluorochrome Information:
Super Bright 645 is a tandem dye excited by the violet laser (405 nm) and emits at 645 nm. Recommended detection filter: 660/20 bandpass. Ensure instrument compatibility.

Staining Considerations:
When using multiple Super Bright dye-conjugated antibodies, use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize non-specific interactions.

Light Sensitivity:
Protect from light to prevent photo-induced oxidation.

Fixation:
Samples may be stored in IC Fixation Buffer (Product # 00-8222) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days at 4°C in the dark with minimal impact on brightness.

Excitation: 405 nm
Emission: 645 nm
Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

Target Information

TIM3 (Hepatitis A virus cellular receptor 2, HAVCR2, T-cell immunoglobulin and mucin-domain containing-3)
TIM3 is a 281 amino acid, type I Th1-specific surface glycoprotein expressed on terminally differentiated CD4⁺ Th1 and CD8⁺ Tc1 cells. It includes an IgV-like domain, a mucin-like extracellular domain, and a conserved tyrosine phosphorylation motif in the cytoplasmic region. TIM3 is involved in macrophage activation, autoimmune disease induction, and regulation of Th1-mediated immune responses, contributing to immune tolerance. It is implicated in experimental autoimmune encephalomyelitis (EAE), enhancing disease severity in mice.

For Research Use Only.
Not for use in diagnostic procedures or resale without authorization.

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