Anti-Mouse TIM-3 (Clone B8.2C12) – Purified in vivo PLATINUM™ Functional Grade
Anti-Mouse TIM-3 (Clone B8.2C12) – Purified in vivo PLATINUM™ Functional Grade
Product No.: T752
Clone B8.2C12 Target Tim-3 Formats AvailableView All Product Type Hybridoma Monoclonal Antibody
Alternate Names HAVCR2 Isotype Rat IgG1 κ Applications FA , FC |
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Dilution Buffer Immunogen Mouse TIM-3 protein Product Concentration ≥ 5.0 mg/ml Endotoxin Level <0.5 EU/mg as determined by the LAL method Purity ≥98% 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. State of Matter Liquid 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. Pathogen Testing To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile. 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. Regulatory Status Research Use Only Country of Origin USA Shipping 2 – 8° C Wet Ice Additional Applications Reported In Literature ? FA, FC Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity B8.2C12 activity is directed against mouse TIM-3 (CD366). Background TIM-3 is a member of the T cell immunoglobulin mucin gene family and encodes a type I
membrane protein consisting of an immunoglobulin variable-region-like domain, a mucin-like
domain, and a tyrosine phosphorylation motif1. TIM-3 functions as an important immune checkpoint receptor that helps regulate dendritic cell function2, T helper type I expansion, and induction of peripheral tolerance3. TIM-3 interacts with GAL-9, PtdSer, HMGB1 and CEACAM1 to activate biochemical pathways such as immune tolerance, T cell depletion, NF-κB signaling, and IL-2 secretion4. Additionally, TIM-3 expression correlates with terminal differentiation and exhaustion in tumors as well as chronic infection3. Due to its dysregulation in different types of cancer, TIM-3 blockade is being investigated as an anti-tumor immunotherapy2,4. TIM-3 also has potential as a prognostic marker in solid tumors4. B8.2C12 was generated by immunizing Lewis and Lou/M female rats (Harlan Sprague-Dawley) with Th1 polarized T cell clones and/or lines, including Th1-specific clone AE7 and in vitro differentiated Th1 cell lines derived from 5B6 and DO11.10 T cell receptor transgenic mice1. Spleen cells were fused with myeloma cells and a large panel of monoclonal antibodies was screened on Th1 and Th2 cells by flow cytometry. B8.2C12 selectively stained Th1 cells. Gene expression cloning was then used to identify a complementary DNA, which was TIM-3. Antigen Distribution TIM-3 (CD366) is expressed on interferon-γ producing T cells, dendritic
cells, cytotoxic lymphocytes, exhausted T cells, natural killer cells, Th17, and myeloid cells.
TIM-3 is also expressed on CD8 + T cells in the tumor microenvironment as well as intratumoral
macrophages and monocytes. TIM-3 is expressed by T helper 1 cells after several rounds of
polarization in vitro. TIM-3 is not expressed by naïve T cells.
Ligand/Receptor Expressed on activated Th1 lymphocytes, CD11b+ macrophages, and dendritic cells. NCBI Gene Bank ID UniProt.org Research Area Cell Biology . Immunology . Immune Checkpoint 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 B8.2C12 is a rat monoclonal antibody against mouse TIM-3 (CD366) that is widely used in murine immunology research, but its binding is specific to the BALB/c allele of TIM-3 (with significantly weaker reactivity to the C57BL/6 allele). The most common in vivo applications in mice for clone B8.2C12 include:
Important technical note:
Summary Table: Common In Vivo Applications of Clone B8.2C12 in Mice
There are other clones (such as RMT3-23) that do not exhibit this strain specificity and may be more appropriate for C57BL/6 mice. When designing experiments, it is essential to ensure BALB/c or compatible strains are used for in vivo studies with B8.2C12. Based on the available information, B8.2C12 is commonly used alongside several other antibodies and proteins in research contexts, particularly in studies involving TIM-3 (CD366) immune checkpoint signaling. Other Anti-TIM-3 Antibody ClonesThe most frequently mentioned companion antibodies to B8.2C12 are RMT3-23 and 5D12, which are different anti-TIM-3 clones used for comparative studies. These three clones differ in their species origin and isotype characteristics: B8.2C12 is a rat IgG1, RMT3-23 is a rat IgG2a, and 5D12 is a mouse IgG1. Notably, while B8.2C12 binds specifically to the BALB/c allele of TIM-3 with significantly weaker reactivity to the C57Bl/6 allele, RMT3-23 can efficiently bind to both BALB/c and C57BL/6 TIM-3. These clones are often used in combination to block different epitopes or compared to evaluate their relative efficacy in functional studies. Co-Checkpoint InhibitorsB8.2C12 is studied in conjunction with other immune checkpoint proteins, particularly PD-1. Research has shown that CD8 T cells expressing both TIM-3 and PD-1 exhibit greater defects in cell-cycle progression and effector cytokine production compared to cells expressing PD-1 alone. This suggests that B8.2C12 may be used in experiments examining dual checkpoint blockade strategies. TIM-3 Ligands and Binding PartnersB8.2C12 is used to study TIM-3 interactions with several key binding partners, including galectin-9 (GAL-9), phosphatidylserine (PtdSer), HMGB1, and CEACAM1. These ligands activate various biochemical pathways such as immune tolerance, T cell depletion, NF-κB signaling, and IL-2 secretion. Key findings from scientific literature on clone B8.2C12 focus on its specificity as an anti-mouse Tim-3 antibody, its restricted allele binding, and its applications in immunological research:
Summary Table: Clone B8.2C12 Findings
These findings highlight B8.2C12’s high allele specificity, utility in immunophenotyping, flow cytometry, and checkpoint blockade research, but also emphasize careful strain selection due to its restricted binding profile. Dosing regimens for clone B8.2C12, an anti-mouse TIM-3 antibody, can vary across different mouse models depending on several factors, including the mouse strain, age, and disease/model system being studied. Here are some general considerations and specific uses: General Considerations for Dosing Regimens
Specific Applications
Dosing Regimen ExampleWhile specific dosing details for B8.2C12 are not widely reported, other antibodies like RMT3-23, another anti-Tim-3 clone, are administered at dosages around 200 µg per mouse via i.p. injection, repeated every few days. The exact dosing for B8.2C12 would require titration to ensure optimal performance for each specific application. References & Citations1 Monney L, Sabatos CA, Gaglia JL, et al. Nature. 415(6871):536-541. 2002. 2 Dixon KO, Tabaka M, Schramm MA, et al. Nature. 595(7865):101-106. 2021. 3 Sabatos CA, Chakravarti S, Cha E, et al. Nat Immunol. 4(11):1102-1110. 2003. 4 Sauer N, Janicka N, Szlasa W, et al. Cancer Immunol Immunother. 72(11):3405-3425. 2023. 5 del Rio ML, Cote-Sierra J, Rodriguez-Barbosa JI. Transpl Int. 24(5):501-513. 2011. 6 Cong J, Wang X, Zheng X, et al. Cell Metab. 28(2):243-255.e5. 2018. 7 Chen L, Yang QC, Li YC, et al. Cancer Immunol Res. 8(2):179-191. 2020. 8 Taniguchi H, Caeser R, Chavan SS, et al. Cell Rep. 39(7):110814. 2022. 9 Guo J, De May H, Franco S, et al. Nat Biomed Eng. 6(1):19-31. 2022. Technical ProtocolsCertificate of Analysis |
Formats Available
Prod No. | Description |
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T751 | |
T756 | |
T757 | |
T758 | |
T752 |
