Anti-Mouse CD366 (Tim-3) [Clone RMT3-23] — Purified in vivo GOLD™ Functional Grade
Anti-Mouse CD366 (Tim-3) [Clone RMT3-23] — Purified in vivo GOLD™ Functional Grade
Product No.: T700
Clone RMT3-23 Target Tim-3 Formats AvailableView All Product Type Monoclonal Antibody Alternate Names Havcr2 Isotype Rat IgG2a κ Applications B , FC , IF Staining , IHC , IHC FF , in vivo , PhenoCycler® |
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Isotype Controls Recommended Dilution Buffer Immunogen This antibody was produced using recombinant mouse TIM-3 Product Concentration ≥ 5.0 mg/ml Endotoxin Level < 1.0 EU/mg as determined by the LAL method Purity ≥95% monomer by analytical SEC ⋅ >95% by SDS Page Formulation This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration. Product Preparation Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates. Storage and Handling Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles. Country of Origin USA Shipping Next Day 2-8°C RRIDAB_2737574 Applications and Recommended Usage? Quality Tested by Leinco FC The suggested concentration for Tim-3 antibody (clone RMT3-23) for staining cells in flow cytometry is ≤ 1.0 μg per 106 cells in a volume of 100 μl or 100μl of whole blood. Titration of the reagent is recommended for optimal performance for each application. Additional Applications Reported In Literature ? B IHC Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Tim-3 antibody (clone RMT2-23) recognizes an epitope on mouse Tim-3. Background Tim-3 antibody (clone RMT3-23) recognizes T cell immunoglobulin and mucin domain-containing protein 3 (TIM3, also known as CD366), a TIM family member of immunoregulatory proteins. Tim-3 is a 60kDa type I transmembrane protein with an extracellular immunoglobulin and mucin-like domain and a cytoplasmic tyrosine phosphorylation motif. Tim-3 is expressed by IFNg-producing Th1 CD4 and Tc1 (cytotoxic) CD8 T cells1, regulatory T cells (Tregs)2, myeloid cells3, NK cells4, and mast cells5. Tim-3 is an inhibitory molecule that limits Th1-mediated inflammatory diseases, including a model of central nervous system inflammation (experimental autoimmune encephalomyelitis, EAE)1, inflammatory bowel disease (IBD)6, and type I diabetes7. Tim-3 is also proposed to induce immunological tolerance7, promote Th1 apoptosis8, and regulate macrophage activation1. In addition, Tim-3 is upregulated on tumor-infiltrating lymphocytes, and coblockade of Tim-3 and immune checkpoint inhibitors, such as PD-1, is currently being investigated in clinical trials for the treatment of cancer9. Antigen Distribution Tim-3 is expressed on activated Th1 and Tc1 lymphocytes, CD11b+ macrophages, Tregs, NK cells, and mast cells.
Ligand/Receptor Putative ligand on resting CD4+ lymphocytes Function May play a role in the development of immune responses and the development of Th1-mediated responses PubMed NCBI Gene Bank ID UniProt.org Research Area Immunology . Inhibitory Molecules Leinco Antibody AdvisorPowered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments. Clone RMT3-23 is a rat monoclonal antibody against mouse TIM-3 (CD366), and is widely used in mice for in vivo functional blockade of the TIM-3 immune checkpoint receptor, particularly to investigate TIM-3’s role in immune regulation, tumor immunity, and autoimmunity. Key in vivo applications include:
RMT3-23 is frequently administered intraperitoneally (i.p.) in these studies at defined doses (e.g., 250–500 µg per injection), and its effects are monitored via flow cytometry, cytokine analysis, and functional immune assays. In summary: Commonly used antibodies and proteins in the literature alongside RMT3-23 (an anti-mouse TIM-3/CD366 antibody) include other immune checkpoint inhibitors such as anti-PD-1 and anti-PD-L1 antibodies, as well as antibodies targeting LAG-3 and alternative anti-TIM-3 clones like B8.2C12. These combinations are frequently used to study synergistic effects on immune modulation, especially in tumor immunity research. Key points:
These combinations are especially important in preclinical tumor models, autoimmune disease studies, and immune exhaustion research, reflecting the central role of RMT3-23 within broader immunoregulatory investigations. Clone RMT3-23 is a widely used rat IgG2a monoclonal antibody targeting mouse Tim-3 (CD366), an immunoregulatory receptor involved in T cell exhaustion and tumor immunity. Key findings from scientific citations using clone RMT3-23 are:
These findings position RMT3-23 as a gold-standard tool for dissecting Tim-3 mediated immunoregulation and for developing combinatorial immunotherapy strategies in preclinical mouse models. Dosing regimens of clone RMT3-23 (anti-TIM-3) in mouse models vary significantly based on the experimental context, disease model, and therapeutic goals. The protocols differ primarily in dose amounts, frequency, and timing of administration. Maternal Tolerance and Pregnancy ModelsIn pregnancy and maternal tolerance studies, RMT3-23 follows a specific declining dose schedule. Pregnant mice receive four intraperitoneal injections at 500 μg, 500 μg, 250 μg, and 250 μg on gestational days 6.5, 8.5, 10.5, and 12.5 respectively. This protocol uses higher initial doses that taper over time, reflecting the critical windows of immune regulation during early pregnancy. This dosing schedule has been well-documented and represents the most robust published protocol for RMT3-23. Cancer Immunotherapy ModelsTumor models employ different dosing strategies compared to pregnancy studies. In checkpoint inhibitor combination therapy experiments using CT26 and MC38 tumor xenografts, mice receive 250 μg of anti-TIM-3 (RMT3-23) on days 3, 6, and 9 following tumor inoculation. This regimen uses consistent doses throughout treatment and is administered in combination with other checkpoint inhibitors like anti-PD-1 (200 μg) as part of combination immunotherapy protocols. The dosing is designed to approximate the 3-10 mg/kg range used in human clinical studies when scaled appropriately for mice. Key Dosing ConsiderationsThe choice of dosing regimen depends on several factors:
The variation in these protocols reflects the distinct immunological mechanisms at play—immune tolerance in pregnancy versus anti-tumor immunity—requiring tailored approaches to effectively modulate TIM-3 signaling in each context. References & Citations1. Monney, L. et al. (2002)Nature 415, 536–541. 2. Gao, X. et al. (2012) PLOS ONE 7, e30676. 3. Anderson, A. C. et al. (2007) Science 318, 1141–1143. 4. Ndhlovu, L. C. et al. (2012) Blood 119, 3734–3743. 5. Phong, B. L. et al. J. (2015) Exp. Med. 212, 2289–2304. 6. Li, X. et al. (2010) Clin. Immunol. 134, 169–177. 7. Sanchez-Fueyo, A. et al. (2003) Nat. Immunol. 4, 1093–1101. 8. Zhu C, et al.. Nature Immunology. 2005;6:1245–1252. Technical ProtocolsCertificate of Analysis |
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