Recombinant Mouse IL-4 Rα

Recombinant Mouse IL-4 Rα is used in research applications to study and manipulate the IL-4/IL-13 signaling axis, which is central to immune regulation, inflammation, and disease modeling. This recombinant protein enables precise control and investigation of IL-4Rα-mediated pathways in vitro and in vivo.

Key scientific applications and rationale include:

• Immune Modulation Studies: IL-4Rα is the shared receptor subunit for both IL-4 and IL-13, mediating type 2 immune responses such as Th2 cell differentiation, alternative macrophage activation, and eosinophil recruitment. Recombinant IL-4 Rα allows researchers to dissect these pathways by acting as a decoy receptor, blocking endogenous ligand-receptor interactions, or by serving as a binding partner in mechanistic assays.

• Disease Modeling: IL-4Rα is implicated in the pathogenesis of asthma, atopic dermatitis, allergic rhinitis, and other autoimmune or inflammatory diseases. Recombinant IL-4 Rα can be used to model these diseases in vitro (e.g., by inhibiting IL-4/IL-13 signaling in cell cultures) or in vivo (e.g., by modulating immune responses in mouse models).

• Functional Assays: The biological activity of recombinant Mouse IL-4 Rα is commonly assessed by its ability to inhibit IL-4-dependent cell proliferation (e.g., HT2 cell assays), making it a valuable tool for quantifying IL-4 bioactivity and screening for antagonists or inhibitors.

• Mechanistic Studies: Recombinant IL-4 Rα is essential for investigating receptor-ligand interactions, downstream signaling events (such as STAT6 activation), and the effects of genetic variants or mutations (e.g., Q576R) on receptor function and disease susceptibility.

• Therapeutic Research: IL-4Rα is a validated therapeutic target for monoclonal antibodies in clinical development for type 2 inflammatory diseases. Recombinant receptor proteins are used in preclinical studies to evaluate drug efficacy, specificity, and mechanism of action.

Best practices for using recombinant Mouse IL-4 Rα include:

• Employing it as a soluble decoy receptor to neutralize IL-4 or IL-13 in cell-based assays.
• Using it in binding studies to characterize ligand specificity and affinity.
• Applying it in animal models to modulate immune responses and study disease mechanisms.

In summary, recombinant Mouse IL-4 Rα is a versatile tool for dissecting immune signaling, modeling disease, and developing therapeutics targeting the IL-4/IL-13 axis. Its use is essential for mechanistic, translational, and therapeutic research in immunology and related fields.

Recombinant Mouse IL-4 Rα (Interleukin-4 Receptor alpha) is generally not suitable as a standard for quantification or calibration in ELISA assays designed to measure IL-4 cytokine levels. ELISA standards must match the analyte being quantified—in this case, mouse IL-4, not its receptor.

Key points:

• ELISA standards for mouse IL-4 quantification are typically recombinant mouse IL-4 protein, not IL-4 Rα. The standard curve is generated using known concentrations of IL-4, allowing accurate quantification of IL-4 in samples.
• IL-4 Rα is the receptor for IL-4, not the cytokine itself. Its structure, immunoreactivity, and molecular weight differ from IL-4, so it cannot be used to calibrate assays for IL-4 quantification.
• Using IL-4 Rα as a standard would result in inaccurate quantification, as the antibodies in IL-4 ELISA kits are specific for IL-4, not its receptor. The assay would not recognize IL-4 Rα in the same way, leading to invalid standard curves and unreliable results.
• Validated protocols and commercial kits universally use recombinant mouse IL-4 as the standard for ELISA calibration.

Best practice:
Always use a recombinant mouse IL-4 protein standard for quantification or calibration in ELISA assays measuring IL-4. If you need to quantify IL-4 Rα (the receptor), you must use an ELISA specifically designed for IL-4 Rα, with appropriate standards and antibodies for that target.

Summary Table:

Do not substitute IL-4 Rα for IL-4 in ELISA calibration. Use the standard that matches your analyte for accurate quantification.

Recombinant Mouse IL-4 Rα has been primarily validated for use in bioassays in published research, particularly for studying cytokine signaling, immune cell differentiation, and functional modulation of immune responses in mouse models.

Key validated applications in published research include:

• Bioassays: The recombinant protein is widely used in cell-based functional assays to study IL-4/IL-13 signaling, T cell and B cell responses, macrophage polarization, and cytokine-induced gene expression.
• In vitro studies: It is used to assess the effects of IL-4Rα engagement or blockade on immune cell proliferation, differentiation (e.g., Th2 polarization), and activation, often in the context of allergic inflammation, tumor immunology, and autoimmune disease models.
• In vivo models: Recombinant mouse IL-4 Rα has been used in mouse models to investigate the role of IL-4/IL-13 signaling in diseases such as asthma, atopic dermatitis, and tumor immunity, often by administering the protein or its antagonists to modulate immune responses.
• Mechanistic studies: Published research has used recombinant IL-4 Rα to dissect signaling pathways, such as STAT6 activation, and to evaluate the effects of receptor engagement on downstream gene expression and cellular phenotypes.

Representative published research applications:

• Tumor immunology: Used in studies examining T cell responses, macrophage polarization, and tumor microenvironment modulation.
• Allergic and autoimmune disease models: Applied in asthma, atopic dermatitis, and other type 2 inflammation models to study the therapeutic potential of IL-4Rα targeting.
• Cellular differentiation: Used to induce or block Th2 differentiation in T cells and to study B cell class switching.

Summary Table:

Note: The majority of published research validates recombinant mouse IL-4 Rα for use in bioassays with whole cells, both in vitro and in vivo, to study immune signaling and disease mechanisms. There is no evidence in the provided results for use in applications such as Western blot, ELISA, or immunohistochemistry; its primary use is functional and mechanistic cell-based assays.

To reconstitute and prepare Recombinant Mouse IL-4 Rα protein for cell culture experiments, follow these general best practices based on manufacturer recommendations and scientific protocols:

1. Reconstitution

• Buffer: Reconstitute the lyophilized protein in sterile, cold phosphate-buffered saline (PBS) or a suitable cell culture-compatible buffer (e.g., sterile PBS with 0.1% carrier protein such as BSA or HSA, if recommended). Some protocols may specify a different buffer, so always check the Certificate of Analysis (CoA) or product datasheet for specific instructions.
• Carrier Protein: If the protein is carrier-free, it is often recommended to add a carrier protein (e.g., 0.1% BSA or HSA) to stabilize the cytokine and prevent adsorption to surfaces.
• Volume: Reconstitute at a concentration recommended by the manufacturer (commonly 100 µg/mL or as specified). For example, for a 10 µg vial, add 100 µL of buffer to achieve 100 µg/mL.
• Procedure: Briefly centrifuge the vial before opening to ensure all powder is at the bottom. Add the buffer slowly and gently mix by pipetting up and down or swirling. Do not vortex, as this can denature the protein. Allow several minutes for complete reconstitution.

2. Preparation for Cell Culture

• Dilution: After reconstitution, dilute the stock solution in complete cell culture medium (containing serum or other carrier proteins) to the desired working concentration. Typical working concentrations for IL-4 Rα in cell culture range from 1–100 ng/mL, depending on the experimental design.
• Sterility: Ensure all steps are performed under sterile conditions to avoid contamination.
• Storage: Store reconstituted protein at 2–8°C for short-term use (up to one month) or aliquot and store at –20°C to –70°C for long-term storage. Avoid repeated freeze-thaw cycles.

3. Additional Tips

• Activity: Confirm the biological activity of the protein using a relevant bioassay (e.g., proliferation assay with a responsive cell line).
• Controls: Include appropriate controls (e.g., untreated cells, vehicle controls) in your experiments.

Always refer to the specific product datasheet or CoA for any manufacturer-specific instructions, as protocols may vary slightly between suppliers.