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

[product_table name="All Top" skus="T700"]

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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®

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
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
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.

Description

Description

Specificity
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
Research Area
Immunology
.
Inhibitory Molecules

Leinco Antibody Advisor

Powered 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:

  • Immune checkpoint blockade for cancer immunotherapy research: RMT3-23 is administered to mice to block TIM-3, releasing inhibition on T cell responses, and is used to examine antitumor immunity, either alone or in combination with other checkpoint inhibitors. This can lead to suppression of tumor cell growth in murine cancer models.

  • Study of autoimmunity and inflammatory diseases: RMT3-23 is applied to assess TIM-3's function as a negative regulator of Th1-mediated diseases (e.g., experimental autoimmune encephalomyelitis for multiple sclerosis models, graft-versus-host disease, type 1 diabetes, and inflammatory bowel disease) by promoting or suppressing specific immune responses.

  • Alloimmunity and transplantation tolerance: In mouse models of transplant (e.g., allogeneic skin or cardiac allografts), RMT3-23 is used to modulate T cell responses, test for enhanced alloimmune activation, and to study immune tolerance mechanisms. RMT3-23 treatment can increase the frequency of effector cytokine-producing T cells and regulate Treg induction in transplant models.

  • Regulation of phagocytosis and innate immunity: RMT3-23 has been used to inhibit TIM-3-mediated phagocytosis of apoptotic cells by Mac1+ peritoneal exudate cells, probing TIM-3’s role on myeloid cells and innate immune tolerance.

  • Investigations of other TIM-3–expressing cell types: TIM-3 is also expressed on regulatory T cells, NK cells, mast cells, and myeloid cells, broadening the application of RMT3-23 to various immunological models in vivo.

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:
The most common in vivo applications of clone RMT3-23 in mice are for functional blockade of TIM-3 in studies of tumor immunity, autoimmunity (including EAE and diabetes), transplantation, and regulation of phagocytosis, helping to dissect TIM-3's role as an inhibitory immune checkpoint in diverse immunological contexts.

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:

  • Anti-PD-1 (e.g., clones like RMP1-14 for mouse), anti-PD-L1, and sometimes anti-LAG-3 antibodies are commonly paired with RMT3-23 in combination immunotherapy or mechanistic studies, based on their roles as distinct but complementary immune checkpoint inhibitors.
  • Alternative anti-TIM-3 clones, such as B8.2C12, are often used in comparative studies to validate findings or assess differences in efficacy and specificity between different clones.
  • Proteins or ligands relevant to the TIM-3 pathway, such as galectin-9, PtdSer (phosphatidylserine), and CEACAM1, are also studied in conjunction with RMT3-23 for understanding signaling mechanisms.

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:

  • Epitope specificity: RMT3-23 binds a unique, non-overlapping epitope on mouse Tim-3 distinct from other anti-Tim-3 clones (such as B8.2C12 and 5D12), facilitating reliable detection and mechanistic studies, even during combination antibody therapy or tracking Tim-3+ cells in treated animals.

  • Receptor antagonism and mechanism: RMT3-23 acts as a TIM-3 receptor antagonist, effectively blocking Tim-3 function without depleting Tim-3+ cells in vivo. Unlike some antibodies that induce target cell depletion via Fc effector functions, RMT3-23 primarily down-modulates Tim-3 surface expression, especially when crosslinking occurs, but does not significantly reduce CD4+ or CD8+ T cell numbers, nor Treg frequencies in the tumor microenvironment.

  • Allelic specificity: RMT3-23 binds both BALB/c and C57BL/6 alleles of mouse Tim-3, in contrast to clone B8.2C12, which binds only BALB/c Tim-3. This makes RMT3-23 particularly versatile for murine immunology studies.

  • Preclinical efficacy: In mouse models of cancer (e.g., WT3 sarcoma), RMT3-23 demonstrates monotherapeutic antitumor efficacy and can enhance antitumor immunity when used alone or in combination with anti-PD-1 therapy.

  • Functional studies: RMT3-23 is used to:

    • Block Tim-3:phosphatidylserine (PS) and Tim-3:CEACAM1 interactions, complicating T cell–APC crosstalk and effecting antitumor immunity.
    • Inhibit Tim-3 mediated T cell trogocytosis, which is a process that limits antitumor responses.
    • Study the dynamics and regulation of Tim-3+ immune cells during experimental therapies and infection processes.
  • Usage and detection: RMT3-23 is compatible with a variety of detection platforms (flow cytometry, in vivo blockade, functional assays), facilitating both mechanistic and therapeutic studies in mouse models.

  • Summary Table

CharacteristicKey Finding
IsotypeRat IgG2a
Species reactivityMouse (BALB/c, C57BL/6 alleles)
EpitopeUnique, non-overlapping on Tim-3
Mode of actionTim-3 receptor antagonist; promotes down-modulation
DepletionDoes not deplete Tim-3+ cells
Preclinical utilityReliable for tumor, infection, and T cell exhaustion studies
Combination therapyEffective combined with anti-PD-1 blockade

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 Models

In 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 Models

Tumor 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 Considerations

The choice of dosing regimen depends on several factors:

  • Disease model: Pregnancy models use higher initial doses with tapering, while tumor models maintain consistent doses
  • Treatment duration: Pregnancy protocols span approximately one week across critical gestational periods, whereas cancer models follow tumor growth dynamics
  • Combination therapy: When used with other checkpoint inhibitors, TIM-3 blockade doses are coordinated with companion antibodies
  • Route of administration: All documented protocols use intraperitoneal injection as the standard delivery method

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 & Citations

1. 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.
B
Flow Cytometry
IF Staining
IHC
IHC FF
in vivo Protocol
PhenoCycler®

Certificate of Analysis

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.