Anti-Mouse TIM-4 – Purified in vivo GOLD™ Functional Grade

The RMT4-53 antibody is a **murine monoclonal antibody that blocks TIM-4**, and it is widely used in in vivo mouse studies to dissect the role of TIM-4 in immune regulation, transplantation, and tumor immunity.**Key applications and experimental protocols include:**- **Blockade of TIM-4 signaling in vivo:** RMT4-53 is administered to mice (typically by intraperitoneal injection) to block TIM-4 function on antigen-presenting cells, thereby altering T cell responses and regulatory T cell (Treg) dynamics[1][3][4][5].- **Transplantation studies:** RMT4-53 has been used to prolong allograft survival. In skin graft models, administration of RMT4-53 leads to enhanced conversion of CD4^+^ T cells into induced Tregs (iTregs) in vivo, reducing effector T cell expansion and promoting tolerance to the graft[1]. Graft-prolonging effects are critically dependent on the capacity for iTreg generation by transferred T cells[1].- **Tumor immunity:** In murine cancer models (such as melanoma and colon adenocarcinoma), RMT4-53 potentiates antitumor immune responses. It enhances the frequency of interferon gamma (IFN-?)-producing CD8^+^ T cells and inhibits tumor growth, especially when combined with vaccine strategies[5].- **Adoptive transfer and mechanistic studies:** RMT4-53 is used in adoptive transfer models to track antigen-specific T cell populations and Treg conversion, typically by analyzing lymphoid organs post-treatment[1].**Mechanism of action:**- By binding to TIM-4, RMT4-53 blocks its interaction with ligands, such as phosphatidylserine, thereby interfering with processes like **apoptotic cell uptake**, **antigen presentation**, and **T cell activation or regulation**[2][3][5].**Experimental details:**- Dosing regimens and administration routes (e.g., dose per mouse, frequency, and method) may vary depending on the experiment, but the antibody is sold as a purified, endotoxin-low reagent suitable for in vivo use[3].- The antibody is often compared to isotype control antibodies to confirm specificity of effects[3].**Summary Table: Main Uses of RMT4-53 in Mouse Studies**| Application Area | Experimental Role | Effect/Outcome ||---------------------|--------------------------------------------------|--------------------------------------------------|| Allograft/transplantation | Inhibits TIM-4 to induce iTregs, prolongs graft survival | Increased Foxp3^+^ iTreg conversion, reduced effector T cells, prolonged survival[1] || Tumor immunity | Blocks TIM-4 during vaccination or tumor challenge | Elevated anti-tumor T cell response, tumor inhibition[5] || Mechanistic T cell studies | Tracks Treg induction and T cell proliferation | Reveals dependence on DCs/TIM-4 for tolerance induction[1] |RMT4-53 is thus a standard tool for studying TIM-4 biology and therapeutics in mouse models of transplantation, tolerance, and cancer[1][3][4][5].

The correct storage temperature for sterile packaged **clone RMT4-53** (Anti-Mouse TIMD4/TIM-4 Antibody) depends on the duration and formulation:- **Short-term storage (1–2 weeks):** Store at **4°C**[1][3][7].- **Medium-term storage (up to 12 months):** Store at **-20°C**[1][3].- **Long-term storage:** Store at **-80°C**[1][3].Some formulations intended for flow cytometry (e.g., conjugated to APC) are specifically labeled for storage at **4°C for up to 12 months, protected from light, and not to be frozen**[5]. Always check the specific product datasheet for formulation-specific recommendations, as preservatives and conjugates may alter storage needs.For standard unconjugated and sterile antibodies, these guidelines above apply broadly and are supported by multiple sources. Avoid repeated freeze-thaw cycles and aliquot the antibody as needed to prevent degradation[4].

RMT4-53, a rat monoclonal antibody targeting **mouse TIM-4 (TIMD4)**, is commonly used in immunological research, often in combination with other antibodies and proteins to dissect immune mechanisms and therapeutic effects.Commonly co-used antibodies or proteins include:- **FoxP3**: For identification and isolation of regulatory T cells (Tregs), particularly when assessing the impact of TIM-4 blockade on Treg induction and function[2].- **CD4 and CD25**: Used to characterize T cell populations, especially when generating or analyzing induced regulatory T cells (iTregs) from naïve CD4^+^CD25^?^ cells[2].- **IFN-? detection antibodies**: Utilized in ELISPOT or Luminex assays to measure effector function of T cells, particularly the production of interferon gamma by CD8^+ or CD4^+ cells following TIM-4 blockade[1][2].- **Isotype controls**: Rat IgG2b isotype control antibodies are recommended for specificity verification in in vivo and in vitro assays when RMT4-53 is used[3].- **Cancer vaccines (e.g., FVAX, DNA vaccines)**: These are not antibodies, but relevant protein reagents often combined with RMT4-53 to study synergistic effects on tumor immunity[1].- **Cytotoxic chemotherapy agents (e.g., cisplatin, oxaliplatin)**: Again not antibodies, but their combination with RMT4-53 is well-documented in tumor models to examine additive effects on anti-tumor immune responses[1].- **Other TIM family antibodies (e.g., RMT3-23 for TIM-3)**: Used in conjunction with RMT4-53 to explore combinatorial blockade of TIM pathways for enhanced immune activation or tumor therapy[1].These combinations allow researchers to:- Assess **immune cell phenotype** and function (e.g., by flow cytometry with CD4, CD25, FoxP3, IFN-?).- Evaluate the **synergistic or additive effects** on immune responses and tumor control when combined with other modulators such as cancer vaccines, checkpoint blockade, or chemotherapeutics[1][2].RMT4-53 is generally used with the above reagents in models involving **mouse immune cells**, especially in studies focused on immune regulation, phagocytosis of apoptotic cells, induction of tolerance, and anti-tumor immunity[1][2][3].

The RMT4-53 clone is a monoclonal antibody targeting TIM4, and its key findings in scientific literature include:1. **Promotion of Islet Graft Survival**: In models of the anti-islet response, RMT4-53 treatment promotes islet graft survival by skewing the immune response from Th1 to Th2. This is observed in Th1-mediated models, where it increases the Th2/Th1 ratio and enhances IL-4 production, indicating a shift towards a more tolerant immune response[1].2. **Effects on Immune Cells**: RMT4-53 treatment decreases the percentage of TIM4-positive B cells and dendritic cells in splenocytes, which are crucial for immune responses. However, it does not significantly alter the percentages of Th17 and CD4^+^ T effector cells but reduces regulatory T cells (Tregs)[1].3. **Tumor Growth Inhibition**: The RMT4-53 clone has been noted to increase the frequencies of IFN-?-producing CD8^+^ T cells, which is associated with the inhibition of tumor growth. This suggests a potential therapeutic role in cancer by enhancing anti-tumor immune responses[3].Overall, RMT4-53 demonstrates potential in modulating immune responses for both transplantation and cancer treatment by targeting TIM4.