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

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

Product No.: T831

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Clone
RMT4-53
Target
TIM-4
Formats AvailableView All
Product Type
Hybridoma Monoclonal Antibody
Alternate Names
T cell immunoglobulin and mucin domain containing protein-4
Isotype
Rat IgG2b κ
Applications
B
,
IF

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Dilution Buffer
Immunogen
Extracellular domains of TIM-4 (aa 1-288)
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.
State of Matter
Liquid
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using only in vitro protein free 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.
Regulatory Status
Research Use Only
Country of Origin
USA
Shipping
2 – 8° C Wet Ice
Additional Applications Reported In Literature ?
B,
IF
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
RMT4-53 activity is directed against mouse TIM-4.
Background
The T cell immunoglobulin and mucin domain containing protein (TIM) family encodes cell surface receptors that are involved in the regulation of T helper (Th) -1 and -2 cell-mediated immunity1. TIM-4, which is preferentially expressed on macrophages and dendritic cells, is the natural ligand of TIM-1, and this binding leads to T-cell expansion and cytokine production. Unlike other members of the TIM family, TIM-4 lacks a putative tyrosine phosphorylation signal sequence in its intracellular domain. The TIM-4 gene maps to a locus associated with predisposition to asthma in both mice and humans and with its connection to TIM-1-triggered Th2 responsiveness, may be considered as a candidate disease/predisposition gene for asthma.

RMT4-53 was generated by linking the extracellular domains of TIM-4 (aa 1-288) to the Fc portion of mouse IgG2a2. This protein product was then used to immunize Sprague Dawley rats. Subsequently, LN cells were fused with P3U1 myeloma cells. RMT4-53 reacts with TIM-4/NRK cells but not with parental NRK or other TIM family members.

TIM-4 blockade has been investigated for the treatment of cancer3,4 and allograft rejection2,5 using anti-TIM-4 clone RMT4-53. Additionally, RMT4-53 blockade of TIM-4 leads to increased induction of iTregs from naïve CD4+ T cells2. Blockade with RMT4-53 has also been investigated in liver ischemia-reperfusion injury6,7.

Antigen Distribution
TIM-4 is expressed by antigen-presenting cells of the lymphoid lineage, preferentially by mature dendritic cells and macrophages.
Ligand/Receptor
Phosphatidylserine, TIM1
NCBI Gene Bank ID
UniProt.org
Research Area
Immunology

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 RMT4-53 is a rat monoclonal antibody that targets mouse TIM-4 and is commonly used in vivo to block TIM-4 signaling in mice. The principal in vivo applications include investigating immune modulation, particularly in transplantation, autoimmunity, and cancer.

Key in vivo applications:

  • TIM-4 Blockade in Immune Regulation Studies:
    RMT4-53 is primarily used to block TIM-4, a receptor found on antigen-presenting cells, to dissect its functional role in modulating immune responses. This application is central to studies investigating how TIM-4 regulates T cell activation, proliferation, and costimulation.

  • Transplantation Models:
    RMT4-53 has been used to study its effect on alloimmune responses in murine transplantation models. For example, in islet transplantation, in vivo targeting of TIM-4 with RMT4-53 led to prolonged islet graft survival and shifted the immune response from a Th1- to a Th2-type (anti-inflammatory). This was associated with reduced splenic TIM-4+ B cells and dendritic cells, suggesting possible depletion or modulation of these populations.

  • Tumor Immunity and Cancer Therapy:
    In cancer research, RMT4-53 is used to block TIM-4 in studies aimed at enhancing anti-tumor immunity, sometimes in combination with other immunotherapies such as anti-PD-1 agents. By targeting TIM-4, researchers seek to disrupt immunosuppressive signaling in the tumor microenvironment.

  • Autoimmunity and Inflammation:
    Studies use RMT4-53 to explore the role of TIM-4 in autoimmune models and inflammation, given its effects on T cell activation and polarization.

  • Cell Depletion and Functional Modulation:
    In vivo administration of RMT4-53 can reduce the number of TIM-4+ immune cells (e.g., B cells and dendritic cells), possibly through antibody-mediated depletion.

Additional technical notes:

  • RMT4-53 is used at function-blocking doses, with dosing regimens tailored to specific experimental protocols (e.g., repeated intraperitoneal injections for transplant models).
  • While the antibody is also used for in vitro assays and immunofluorescence, its major cited in vivo use is blocking TIM-4 function to study its physiological roles.

In summary, the most common in vivo applications of clone RMT4-53 in mice are for blocking TIM-4 to study immune mechanisms in transplantation tolerance, tumor immunity, and autoimmunity, often by altering antigen-presenting cell and T cell functions.

Commonly used antibodies or proteins with RMT4-53 in the literature include other monoclonal antibodies targeting the TIM family (such as anti-TIM-1, anti-TIM-2, and anti-TIM-3), anti-PD-1, isotype controls, and functional proteins involved in immunological assays.

Key antibodies and proteins frequently used alongside RMT4-53:

  • Anti-TIM-1 antibody (clone RMT1-17): Used for comparative or combination studies addressing distinct members of the TIM family.
  • Anti-TIM-2 antibodies (clones RMT2-14 and RMT2-26): Frequently selected to dissect the overlapping and non-overlapping roles of TIM proteins in immune regulation.
  • Anti-TIM-3 antibody (clone RMT3-23): Included when broader TIM family modulation is investigated.
  • Anti-PD-1 antibody (clone Rmp1-14): Co-administered in checkpoint inhibition and tumor immunology studies, often to assess synergistic effects with TIM-4 blockade.
  • Isotype controls: Typically, rat IgG2b isotype control or anti-keyhole limpet hemocyanin controls are used to ensure specificity of the antibody effect in experimental settings.
  • TGFβ (Transforming Growth Factor Beta): Used in the induction of regulatory T cells (iTregs) in functional assays testing the effect of TIM-4 blockade.
  • Antigenic proteins or fusion constructs: For example, bm12 skin or splenocyte antigens in alloantigen-specific Treg suppression assays, and mouse TIM-4-Ig fusion protein as immunogen for generating antibodies.

Experimental models often also employ:

  • Thy1.1 and Thy1.2 surface marker antibodies: For congenic tracking in adoptive transfer and immune tolerance studies.
  • Foxp3-GFP reporter mice or protein constructs: For identifying and sorting regulatory T cell populations in mechanistic studies on immune tolerance.

In summary, the literature commonly utilizes a combination of TIM family antibodies (TIM-1, TIM-2, TIM-3), checkpoint inhibitors (anti-PD-1), isotype controls, and proteins for regulatory T cell induction and immune cell tracking in conjunction with RMT4-53 for comprehensive analysis of immune regulation and therapeutic strategies.

Key findings from clone RMT4-53, as cited in scientific literature, indicate that it is a monoclonal antibody specifically targeting mouse TIM-4 (T cell immunoglobulin and mucin domain 4), with broad relevance in transplantation immunology and tumor immunity research.

Major findings:

  • Alloimmune Response in Islet Transplantation:
    RMT4-53 administration in a murine model promotes islet graft survival by skewing the immune response toward a Th2 phenotype (increased Th2/Th1 ratio). Key immunological effects include:

    • Decreased TIM-4+ B cells and dendritic cells post-transplantation.
    • Increase in IL-4-producing (Th2) cells, but not IFN-γ-producing (Th1) cells.
    • No significant changes in Th17 or effector CD4+ T cells, but a reduction in Treg percentages in treated mice.
    • In Th2-biased settings (Tbet-deficient mice), RMT4-53 unexpectedly accelerated islet rejection, suggesting context-dependent effects of TIM-4 blockade.
    • In B-cell-depleted mice, RMT4-53 prompted islet rejection, highlighting B cell involvement in TIM-4-mediated tolerance.
  • Immune Modulation in Tumor Contexts:
    RMT4-53 is used as a benchmark for mouse TIM-4 blockade in characterizing new antibodies and investigating TIM-4 function in tumor immunity. In these settings:

    • RMT4-53 binds tightly to mouse TIM-4, with no cross-reactivity for human TIM-4; this specificity is often used as a control.
    • TIM-4 blockade, using RMT4-53 or analogous antibodies, may enhance antitumor immunity and synergize with immune checkpoint inhibitors, although most combination studies now employ more translational antibodies.
    • In mouse xenograft models, TIM-4 blockade does not significantly alter myeloid or T lymphocyte composition in the spleen, supporting a primary effect on antigen presentation and phagocytosis rather than broad lymphocyte depletion.
  • Mechanistic Insights:

    • TIM-4 is a phosphatidylserine receptor, involved in apoptotic cell recognition and clearance, primarily expressed by myeloid cells including dendritic cells.
    • RMT4-53’s targeting of TIM-4 leads to immunomodulation largely through changing myeloid activity and antigen processing, with downstream effects on adaptive immune skewing and graft outcomes.
    • RMT4-53 is species-specific and does not bind human TIM-4, meaning its findings are limited to mouse models but define key mechanisms relevant for downstream human antibody development and translational research.

In summary, clone RMT4-53 is extensively used to demonstrate the role of TIM-4 in immune regulation, particularly in transplantation tolerance, Th2 polarization, and myeloid cell function in mice. It is a critical tool for dissecting myeloid-mediated immune modulation and informs the design of next-generation anti-TIM-4 therapeutics for both transplantation and tumor immunology.

Dosing regimens of clone RMT4-53 (anti-mouse TIM-4 antibody) vary depending on the mouse model and disease context, with most published regimens involving repeat intraperitoneal or intravenous injections at defined intervals and doses tailored to the specific experimental setup.

Key Dosing Schedules Across Mouse Models:

  • Th1-Mediated Islet Transplant Model (C57BL/6 mice with BALB/c islet graft):

    • 500 μg intraperitoneal (i.p.) on day 0
    • 250 μg i.p. on days 2, 4, 6, 8, and 10
    • This regimen significantly prolonged graft survival and reduced TIM-4+ B cells and dendritic cells.
  • Th2 (and Th17) Response Model (Tbet^−/−​ C57BL/6 mice with BALB/c islets):

    • Apparent use of a similar schedule, but the antibody in this context accelerated graft rejection rather than prolonging survival.
  • B-cell–Depleted Recipients:

    • RMT4-53 treatment (same general dosing as above) promoted islet rejection in B-cell–depleted C57BL/6 mice, showing model-specific effects.
  • Liver Ischemia Model:

    • 0.25 mg (250 μg) per mouse intravenously (i.v.) administered either 48 hours or 2 hours before ischemia.
  • Tumor Immunotherapy / Combination Studies:

    • 200 μg per injection, intraperitoneally, every 4 days for three doses in combination experiments with other agents.
  • Skin Graft Models:

    • Although dosing is not always specified, RMT4-53 has been used to promote regulatory T cell induction in vivo; details for these models can sometimes mirror those above or be adjusted by investigators.

Summary Table:

Mouse Model / ContextDoseRouteFrequency/TimingReported Effect
Th1 anti-islet alloresponse500 μg day 0, 250 μg (d2,4,6,8,10)i.p.Multiple, 6 total doses over 10 daysProlonged graft survival
Th2/Th17 response, Tbet^−/−^ miceAs above (implied)i.p.As aboveAccelerated rejection
B-cell–depleted recipientsAs above (implied)i.p.As abovePromoted rejection
Liver ischemia250 μgi.v.Once, 2 h or 48 h before ischemiaModulation of immune response
Tumor models/combination (general)200 μgi.p.Every 4th day × 3Functional blockade, synergy with immunotherapy

Important Context and Considerations:

  • The total dose, intervals, and route of administration (i.p. vs i.v.) are adjusted based on the disease model, anticipated mechanism of action, and pharmacokinetics in the mouse strain used.
  • RMT4-53 has variable effects depending on the immunological context (e.g., prolongs graft survival in Th1 settings but accelerates rejection in Th2).
  • In transplantation and tumor models, repeated dosing (rather than single bolus) is typical to maintain blockade during critical phases of immune response.
  • Reports sometimes extrapolate between models; precise dose and schedule optimization requires consideration of the specific experimental objectives and controls.

If you need the dosing regimen for a particular disease area or genetically modified mouse line not covered above, specifying that context may help refine the answer.

References & Citations

1 Meyers JH, Chakravarti S, Schlesinger D, et al. Nat Immunol. 6(5):455-464. 2005.
2 Yeung MY, McGrath MM, Nakayama M, et al. J Immunol. 191(8):4447-4455. 2013.
3 Baghdadi M, Nagao H, Yoshiyama H, et al. Cancer Immunol Immunother. 62(4):629-637.2013.
4 Ding Q, Mohib K, Kuchroo VK, et al. J Immunol. 199(7):2585-2595. 2017.
5 Vergani A, Gatti F, Lee KM, et al. Cell Transplant. 24(8):1599-1614. 2015.
6 Ji H, Liu Y, Zhang Y, et al. Hepatology. 60(6):2052-2064. 2014.
7 Li J, Zhao X, Liu X, et al. Mol Immunol. 66(2):117-125. 2015.

Formats Available

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