Staining of non-transfected (left) and mouse Tim-3 transfected (right) CHO cells with 0.125 μg of Purified Rat IgG1 Iso Cntrl (cat. 14-4301) (open histogram) or 0.125 μg of Purified 8B.2C12 (colored histogram) followed by FITC Anti-Rat IgG (cat. 11-4811). Total cells were used for analysis.
Contents: Affinity Purified anti-mouse TIM-3 (TIM3, Th1 specific marker, blocking / neutralizing) Catalog Number: 14-5871 Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, 0.09% NaN3 Clone: 8B.2C12 Host/Isotype: Rat IgG1, κ Storage Conditions: Store at 4°C. Avoid repeated freeze/thaw cycles.
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Cat. No.
Size
Price
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14-5871-81
50 ug
14-5871-82
100 ug
14-5871-85
500 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.
Description
The 8B.2C12 monoclonal antibody reacts with mouse Tim-3, a Th1-specific cell surface protein. Tim-3, 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. Tim-3 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 naïve T cells, B cells, macrophages and dendritic cells, do not express Tim-3, at least at the protein level. Tim-3 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, Tim-3 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 Tim-3 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 Tim-3 during T cell activation results in the expansion and activation of macrophages and increased severity of an 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 Tim-3 BALB/c allele. Reactivity to the C57/Bl6 is significantly weaker than BALB/c.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The 8B.2C12 antibody has been reported for use in flow cytometric analysis, and immunoprecipitation. It has also been reported for use in functional assays. (Please use Functional Grade purified 8B.2C12, cat. 16-5871, in functional assays.)
Applications Tested
The 8B.2C12 antibody has been tested by flow cytometric analysis of mouse splenocytes and mouse Tim-3 transfected cells. This can be used at less than or equal to 0.25 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Monney, L., C.A. Sabatos, et al. 2002. Th1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease. Nature 415(6871): 536-41.
McIntire J.J., S.E. Umetsu, et al. 2001. Identification of Tapr (an airway hyperreactivity regulatory locus) and the linked Tim gene family. Nat Immunol. 2(12):1109-16.
Beauregard C, Stevens C, Mayhew E, Niederkorn JY. Cutting edge: atopy promotes Th2 responses to alloantigens and increases the incidence and tempo of corneal allograft rejection. J Immunol. 2005 Jun 1;174(11):6577-81.
(IHC, PubMed)
Frisancho-Kiss S, Nyland JF, Davis SE, Barrett MA, Gatewood SJ, Njoku DB, Cihakova D, Silbergeld EK, Rose NR, Fairweather D.Cutting Edge: T Cell Ig Mucin-3 Reduces Inflammatory Heart Disease by Increasing CTLA-4 during Innate Immunity. J Immunol. 2006 Jun 1;176(11):6411-6415. (FA, PubMed)
