Recombinant Mouse IL-1ra

Recombinant Mouse IL-1ra (interleukin-1 receptor antagonist) is widely used in research to specifically inhibit IL-1 signaling, enabling the study and modulation of inflammatory pathways in mouse models. Its use is critical for dissecting the role of IL-1 in inflammation, autoimmunity, tissue repair, and infection.

Key reasons to use recombinant mouse IL-1ra in research applications:

• Selective IL-1 Pathway Inhibition: IL-1ra binds to the IL-1 receptor without activating it, thereby blocking the pro-inflammatory effects of both IL-1α and IL-1β. This allows researchers to specifically assess the contribution of IL-1 signaling in various disease models.

• Anti-Inflammatory Effects: Administration of recombinant IL-1ra reduces inflammation in a range of mouse models, including rheumatoid arthritis, wound healing, and systemic inflammatory responses. For example, it has been shown to suppress cytokine-induced catabolism in cartilage and reduce joint destruction in arthritis models.

• Therapeutic Modeling: Recombinant mouse IL-1ra is used to model and evaluate potential therapies for inflammatory and autoimmune diseases, such as rheumatoid arthritis, macrophage activation syndrome, and severe infections. Its efficacy in reducing inflammation and tissue damage provides a basis for translational studies.

• Tissue Protection and Repair: IL-1ra promotes a pro-healing microenvironment in tissue injury models, such as diabetic wound healing, by reducing pro-inflammatory cytokines and senescent fibroblasts while increasing anti-inflammatory mediators and growth factors.

• Safety and Immune Regulation: Endogenous and exogenous IL-1ra protect mice from uncontrolled systemic inflammation and cytokine release syndromes, making it a valuable tool for studying immune regulation and adverse event mitigation in preclinical models.

• Mechanistic Studies: Using recombinant mouse IL-1ra enables precise mechanistic studies of IL-1-driven processes, including apoptosis, immune cell recruitment, and cytokine cascades, without off-target effects on other cytokine pathways.

• Species-Specificity: Recombinant mouse IL-1ra is species-matched, ensuring optimal receptor binding and biological activity in mouse models, which is essential for accurate in vivo studies.

In summary, recombinant mouse IL-1ra is a powerful reagent for dissecting IL-1-mediated mechanisms, evaluating anti-inflammatory strategies, and modeling therapeutic interventions in mouse research. Its use is supported by extensive preclinical data demonstrating its efficacy in modulating inflammation, promoting tissue repair, and preventing immune-mediated pathology.

You can use recombinant mouse IL-1ra as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay’s requirements. Most commercial mouse IL-1ra ELISA kits use recombinant mouse IL-1ra as the standard to generate the calibration curve for quantification.

Key considerations:

• Source and Validation: The recombinant IL-1ra should be of high purity and its concentration accurately determined. It must be compatible with the antibodies used in your ELISA, as some antibodies may have different affinities for recombinant versus native protein.
• Standard Curve Preparation: Prepare a fresh standard curve for each assay using serial dilutions of the recombinant IL-1ra in the same matrix as your samples (e.g., serum, plasma, or culture supernatant) to account for potential matrix effects.
• Kit Compatibility: If you are using a commercial ELISA kit, use the standard provided with the kit when possible, as it is optimized for that assay. If substituting with your own recombinant IL-1ra, ensure it is functionally equivalent to the kit standard and validated for use in your specific assay format.
• Documentation: Refer to your ELISA kit’s manual for specific recommendations regarding standard protein source and preparation. Some kits specify that only their provided standard should be used for accurate quantification.

Summary Table: Use of Recombinant Mouse IL-1ra as ELISA Standard

In conclusion: Recombinant mouse IL-1ra is widely used as a standard in quantitative ELISA assays for IL-1ra, but ensure it is validated for your assay system and matches the requirements of your antibodies and detection method.

Recombinant Mouse IL-1ra has been validated for a range of applications in published research, primarily focused on its role as an anti-inflammatory agent in both in vitro and in vivo models. Key validated applications include:

• Blocking IL-1-induced inflammatory responses: Recombinant Mouse IL-1ra is widely used to inhibit the biological activity of IL-1α and IL-1β in cell-based assays, such as suppressing the proliferation of IL-1-responsive cell lines (e.g., D10.G4.1 cells).
• Preclinical therapeutic studies: It has been validated in mouse models for investigating potential treatments for diseases such as rheumatoid arthritis, sepsis, chronic myelogenous leukemia, cystic fibrosis, and gouty arthritis.
• Synergistic immunomodulation: Studies have combined recombinant IL-1ra with other cell therapies (e.g., M2 macrophages derived from mesenchymal stem cells) to enhance anti-inflammatory effects in autoimmune disease models.
• Gene therapy research: Recombinant IL-1ra has been used as a transgene in gene therapy approaches, such as intra-articular delivery for osteoarthritis, demonstrating safety and sustained local expression in clinical trials.
• Inflammasome regulation: It has been validated for studies on inflammasome-mediated inflammation, particularly in cystic fibrosis mouse models, where it reduces neutrophil recruitment, lung damage, and improves survival after infection.
• Gout and crystal-induced inflammation: Recombinant IL-1ra has been used in mouse models of monosodium urate (MSU) crystal-induced peritonitis to study its efficacy in blocking IL-1β-driven inflammation, relevant to gout research.

Additional validated uses include:

• Comparative immunology research and as a reagent in mouse model studies for evaluating anti-inflammatory mechanisms and therapeutic interventions.
• Cell signaling and cytokine antagonism studies, where it serves as a control or experimental agent to dissect IL-1 pathway biology.

These applications are supported by both preclinical and clinical research, demonstrating the protein’s utility in mechanistic studies, therapeutic evaluation, and translational research targeting IL-1-mediated pathologies.

To reconstitute and prepare Recombinant Mouse IL-1ra protein for cell culture experiments, centrifuge the vial briefly to collect the lyophilized powder, then add sterile buffer according to the recommended concentration and protocol. The most common reconstitution is in sterile PBS (pH 7.2–7.4) or sterile distilled water, at a concentration between 0.1–0.5 mg/mL.

Step-by-step protocol:

1. Centrifuge the vial briefly to ensure all powder is at the bottom before opening.
2. Add sterile buffer:
   • For carrier-free protein, use sterile PBS (pH 7.2–7.4) or sterile distilled water.
   • For formulations with carrier protein, use sterile PBS containing at least 0.1% BSA (bovine serum albumin) if recommended.
3. Reconstitution concentration: Prepare at 0.1–0.5 mg/mL for stock solutions. For example, add 1 mL buffer to 0.1–0.5 mg protein.
4. Gently mix by pipetting up and down or swirling. Avoid vigorous vortexing to prevent protein denaturation.
5. Incubate at room temperature for 5–10 minutes to ensure complete dissolution.
6. Aliquot the stock solution to avoid repeated freeze-thaw cycles.
7. Storage:
   • Store aliquots at 2–8°C for up to 1 week.
   • For longer-term storage, keep at –20°C or below.
   • Avoid repeated freeze-thaw cycles to preserve activity.

Preparation for cell culture:

• Dilute the stock solution in cell culture medium to the desired working concentration (typically in the ng/mL range, depending on experimental design and cell type).
• If using carrier-free protein, consider adding a small amount of carrier protein (e.g., 0.1% BSA) to working solutions to minimize adsorption to plasticware, unless BSA interferes with your assay.
• Filter sterilize the final working solution if necessary, using a 0.2 μm filter.

Additional notes:

• Always consult the specific product datasheet for any unique requirements regarding buffer composition or reconstitution procedure.
• For functional assays, typical ED50 values for IL-1ra activity are in the range of 2.5–60 ng/mL, depending on the assay system and cell type.
• If the protein is supplied in a buffer (e.g., PBS), simply dilute as needed for your experiment.

Summary Table:

This protocol ensures optimal solubility, stability, and biological activity of recombinant mouse IL-1ra for cell culture experiments.