Anti-Mouse CD252 (OX40L) [Clone RM134L] – Purified in vivo PLATINUM™ Functional Grade

The RM134L clone is a rat monoclonal antibody that targets mouse OX40L (CD252/CD134L) and is widely used in in vivo mouse studies for blocking OX40/OX40L signaling pathways. This antibody serves as a powerful research tool to investigate the functional role of the OX40-OX40L costimulatory interaction in immune responses.

Primary In Vivo Applications

Immune Response Modulation: The RM134L antibody effectively blocks the costimulatory activity of OX40L, which is crucial for T cell activation and proliferation. When administered in vivo, it disrupts the interaction between OX40L (expressed on antigen-presenting cells and activated B cells) and OX40 (CD134, found on activated T cells).

T Cell Proliferation Studies: In vivo treatment with RM134L has been specifically demonstrated to inhibit poly(I:C)/CD40 stimulated proliferation of CD4 T cells. This makes it particularly valuable for studying T cell responses in various experimental models where researchers need to understand the contribution of OX40/OX40L signaling to immune activation.

Mechanism of Action in Vivo

The RM134L antibody works by neutralizing OX40L function through direct binding, preventing the natural ligand-receptor interaction between OX40L and OX40. This blocking activity is reciprocally significant because the OX40L-OX40 interaction normally provides costimulatory signals that enhance both T cell and B cell responses - stimulation via OX40 increases T cell proliferation and IL-2 production, while OX40L stimulation enhances B cell proliferation and immunoglobulin secretion.

Research Applications

The antibody is particularly useful in studies investigating immune tolerance, autoimmunity, and transplant rejection, where researchers need to modulate T cell costimulation. The availability of endotoxin-free, azide-free formulations (LEAF™ purified) makes it especially suitable for functional in vivo assays where contamination could interfere with experimental results.

The RM134L clone has been extensively validated across multiple research applications and is available from several suppliers in formats optimized for in vivo use, typically as purified, functional-grade antibodies suitable for blocking studies in mouse models.

The correct storage temperature for the sterile packaged clone RM134L (CD252/OX40 Ligand monoclonal antibody) is 4°C, stored in the dark, and DO NOT FREEZE. This guidance is consistent across multiple reputable antibody suppliers.

The packaging and handling instructions specify:

• Temperature: 4°C (standard refrigerator temperature)
• Light protection: Store in the dark to prevent degradation
• Freezing: Do not freeze, as this may compromise antibody integrity

For optimal stability and performance, always follow these conditions unless a specific alternative protocol is indicated on the actual vial or by your supplier.

In the literature, the RM134L antibody, which targets the OX40 Ligand (OX40L or CD252), is often used in conjunction with other antibodies or proteins to study immune responses. Here are some commonly used antibodies or proteins:

1. Anti-IgM and Anti-CD40 Antibodies: These are used to stimulate splenic B cells, which then express OX40L. The combination of these antibodies with RM134L helps in studying the interaction between B cells and T cells, particularly in the context of humoral responses.

2. OX40-Ig Fusion Protein: This protein is used to study the interaction between OX40L and its receptor, OX40. It can bind to OX40L, and its binding is inhibited by the RM134L antibody.

3. Anti-CD3 Antibody: This antibody is used to stimulate T cells, which can be modulated by the interaction between OX40 and OX40L. The RM134L antibody can inhibit the costimulatory activity of OX40L on anti-CD3-stimulated T cells.

4. Lipopolysaccharide (LPS): LPS is used to activate B cells, which may express higher levels of OX40L, although this effect is not consistently detectable with the RM134L antibody.

These reagents are used in various immunological studies, including flow cytometry, to analyze the role of OX40L in immune responses and therapeutic interventions.

Clone RM134L is a monoclonal antibody targeting mouse OX40L (CD252/CD134L), widely used for immunological studies, particularly in mice. Key findings from scientific literature citing clone RM134L are:

• Validated Applications: RM134L is well-established for use in flow cytometry (FC) and immunohistochemistry (IHC) to detect OX40L levels on immune cells, with multiple studies citing its reliability and specificity.

• Role in Immunopathology: RM134L has been crucial in demonstrating the pathogenic role of OX40L in disease models.

  • In a murine model of asthma, OX40L signaling was found to be critical for the development of pathogenic Th2 cells, implicating OX40L as a potential therapeutic target for allergic inflammation.
  • In viral myocarditis (CVB3 infection), anti-OX40L blockade (using RM134L) significantly reduced heart inflammation at later stages of disease in mice, even if it did not conclusively impact early mortality. This suggests OX40L blockade can modulate chronic inflammation after acute viral injury.

• Functional Insights:

  • OX40L blockade using RM134L reduces progression of chronic inflammatory responses in some autoimmune and infectious disease models, demonstrating its utility in dissecting the T cell co-stimulatory pathways during immune activation.
  • OX40L interacts with OX40 (CD134), regulating both CD4+ and CD8+ T cell clonal expansion and effector function, notably on activated B cells and antigen-presenting cells, highlighting the antibody's importance in deciphering costimulatory signaling.

• Scientific References: RM134L has been specifically cited in foundational immunology research, including studies with PMID 12672051, suggesting its impact in understanding Th2 pathology and OX40L’s immunoregulatory functions.

• Species Specificity: RM134L is selective for murine (mouse) OX40L, supporting its use in rodent models but not in human studies.

• Antibody format and storage: It is supplied as a purified rat monoclonal IgG2b?, stable for up to one year at 2–8°C, with distinct lot traceability often required for experimental reproducibility.

Summary Table: Key Findings Using RM134L

RM134L remains a key reagent for studying OX40L biology, T cell co-stimulation, and immune-mediated disease mechanisms in mice.