Anti-Mouse/Human IL-5 – Purified in vivo GOLD™ Functional Grade

Clone TRFK5 is a rat monoclonal antibody used in in vivo mouse studies primarily for neutralizing interleukin-5 (IL-5) and depleting eosinophils. Researchers administer TRFK5 to mice to block endogenous IL-5 activity, which helps investigate the role of IL-5 in immune responses, particularly in models of allergic diseases, asthma, and eosinophil-mediated inflammation.

Key details on in vivo use:

• IL-5 Neutralization: TRFK5 binds to mouse IL-5 and prevents it from interacting with its receptor, effectively neutralizing the cytokine’s biological activity in living animals. This allows scientists to dissect the contribution of IL-5 to B cell differentiation, immunoglobulin production, and particularly eosinophil activation and survival.
• Eosinophil Depletion: By neutralizing IL-5, TRFK5 impairs the survival and maturation of eosinophils, leading to selective eosinophil depletion in vivo. This is a common approach in mouse models of asthma to study the pathophysiology of eosinophilic airway inflammation and allergic responses.
• Typical Administration: The antibody is injected (usually intraperitoneally or intravenously) in mice prior to or during disease induction protocols. Dosing and timing depend on the study design and desired duration of IL-5 blockade.

Other applications include validating the specificity of IL-5-dependent biological responses and serving as an experimental control for anti-eosinophil therapies.

TRFK5 is also widely used for in vitro and ex vivo applications (ELISA, flow cytometry) but its defining role in vivo is as a functional neutralizing and depleting agent for IL-5 and eosinophils in mouse models.

For sterile packaged rat anti-mouse/human IL-5 antibody clone TRFK5, the recommended storage temperature is 2–8°C for regular use. This is in line with storage instructions for both Southern Biotech and BioLegend TRFK5 products, which specify storage of the undiluted antibody in this temperature range.

If long-term storage is required, the antibody should be aseptically aliquoted into working volumes (to avoid repeated freeze-thaw cycles) and stored at –80°C. Some manufacturers also suggest –20°C as an alternative for long-term storage. However, frost-free freezers are not recommended, as temperature fluctuations may compromise antibody integrity.

Summary Table

Always check the specific product datasheet for variations, but these guidelines are widely applicable for TRFK5 monoclonal antibodies.

Based on the literature, TRFK5 is commonly used in conjunction with several other antibodies and proteins in research applications, particularly for IL-5 detection and functional studies.

Complementary Detection Antibodies

TRFK4 is the most frequently paired antibody with TRFK5 in immunoassays. TRFK5 serves as the capture antibody while TRFK4 functions as the detection antibody in sandwich ELISA, ELISpot, and FluoroSpot assays. This pairing creates a comprehensive detection system for mouse IL-5, with TRFK4 often

TRFK5 is a rat monoclonal antibody (clone TRFK5, IgG1 isotype) that specifically targets and neutralizes mouse interleukin-5 (IL-5), and has been extensively used in scientific research to investigate the biological functions of IL-5 in various disease models. The key findings from TRFK5 citations in scientific literature reveal important insights into IL-5's role in cancer progression, wound healing, and immune regulation.

Cancer and Malignant Pleural Effusion Studies

The most significant findings involve TRFK5's therapeutic potential in cancer treatment, particularly in malignant pleural effusion (MPE). Research demonstrated that TRFK5 treatment significantly reduced MPE formation and intrapleural tumor growth in mouse models using Lewis lung cancer (LLC) cells. The antibody showed efficacy in both prevention protocols (administered on days 0 and 8 after tumor cell injection) and regression studies (administered on day 8 after established MPE formation), indicating potential therapeutic benefits even after tumor establishment.

Mechanistic insights revealed that IL-5 neutralization by TRFK5 enhanced pleural tumor cell apoptosis while reducing tumor cell survival. The treatment significantly decreased numbers of eosinophils in both MPE fluid and pleural tumor tissue, suggesting that eosinophil recruitment contributes to IL-5's tumor-promoting effects. Additionally, TRFK5 treatment resulted in significantly lower numbers of CD11b+GR1+ myeloid suppressor cells in MPE fluid compared to controls, indicating that IL-5 promotes tumor growth partly through enhanced recruitment of immunosuppressive cell populations.

Wound Healing Research

TRFK5 has provided crucial insights into eosinophil function in tissue repair processes. In hamster cutaneous wound healing studies, TRFK5 treatment effectively depleted wound-associated eosinophils to levels comparable to unwounded skin. Remarkably, this eosinophil depletion accelerated wound closure by re-epithelialization by 4 days compared to control groups, suggesting that eosinophils may negatively impact certain aspects of wound healing.

The density of eosinophils in day-9 wounds was significantly lower in TRFK5-treated groups, demonstrating the antibody's effectiveness in selectively reducing eosinophil infiltration into healing tissues. This finding challenged previous assumptions about eosinophils' beneficial role in wound repair and highlighted the complex nature of inflammatory cell functions in tissue healing.

Technical Applications and Specifications

TRFK5 has proven versatile across multiple experimental applications. The antibody effectively neutralizes IL-5 bioactivity in functional assays and has been validated for use in ELISA, flow cytometry (requiring membrane permeabilization), and Western blotting applications. Its specificity for mouse IL-5 and lack of cross-reactivity with human, pig, or guinea pig IL-5 makes it particularly valuable for mouse model studies.

The antibody is derived from purified natural murine IL-5 immunization and is prepared by affinity chromatography on Protein A from ascites, ensuring high purity and consistency for research applications. These technical characteristics have made TRFK5 a reliable tool for investigating IL-5 biology across diverse experimental contexts, from cancer research to wound healing and inflammatory disease models.