Recombinant Human IL-4 Receptor Alpha (IL-4 Rα)

Recombinant Human IL-4 Receptor Alpha (IL-4 Rα) is a valuable tool for a wide range of research applications due to its central role in immune regulation and disease pathogenesis. Here are key reasons to use it in your research:

1. Study of Type 2 Immune Responses

IL-4 Rα is a critical component of the receptors for both IL-4 and IL-13, two cytokines that drive type 2 immune responses. These responses are central to allergic diseases (e.g., asthma, atopic dermatitis, allergic rhinitis), parasitic infections, and certain autoimmune conditions. Using recombinant IL-4 Rα allows you to:

• Investigate the molecular mechanisms of IL-4 and IL-13 signaling.
• Study Th2 cell differentiation and function.
• Explore alternative macrophage activation and its implications in immunity and disease.

2. Drug Discovery and Therapeutic Development

IL-4 Rα is a validated therapeutic target, with monoclonal antibodies like dupilumab already approved for clinical use. Recombinant IL-4 Rα is essential for:

• Screening and characterizing novel antagonists, antibodies, or small molecules targeting IL-4/IL-13 signaling.
• Performing binding assays to assess affinity and specificity of candidate therapeutics.
• Validating the mechanism of action of drugs designed to block IL-4 Rα.

3. Functional Assays and Pathway Analysis

Recombinant IL-4 Rα can be used in various functional assays to:

• Measure inhibition of IL-4 or IL-13 signaling in cell-based reporter systems (e.g., HEK-Blue IL-4/IL-13 cells).
• Study downstream signaling events, such as STAT6 phosphorylation.
• Assess the impact of genetic variants or mutations on receptor function.

4. Investigation of Disease Mechanisms

IL-4 Rα is implicated in a broad spectrum of diseases, including:

• Allergic and inflammatory disorders (asthma, atopic dermatitis, allergic rhinitis).
• Autoimmune diseases (rheumatoid arthritis).
• Viral infections (e.g., rhinovirus susceptibility).
• Cancer (e.g., biliary tract cancer, where IL-4 Rα-targeted therapies are being explored).Recombinant IL-4 Rα enables you to model these disease processes and test potential interventions.

5. Protein-Protein Interaction Studies

IL-4 Rα interacts with multiple partners, including IL-4, IL-13, and other receptor subunits. Recombinant IL-4 Rα is ideal for:

• Mapping interaction sites and epitopes.
• Studying the effects of genetic polymorphisms on protein interactions.
• Developing and validating diagnostic tools.

6. Standardization and Quality Control

Recombinant IL-4 Rα provides a consistent, well-characterized reagent for:

• Standardizing assays across different laboratories.
• Ensuring reproducibility in experiments.
• Serving as a positive control in binding and functional assays.

7. Support for Preclinical and Translational Research

Recombinant IL-4 Rα is widely used in preclinical studies to:

• Evaluate the efficacy of novel therapeutics in animal models.
• Support the development of biomarkers for disease diagnosis and monitoring.
• Facilitate the translation of basic research findings into clinical applications.

In summary, recombinant Human IL-4 Receptor Alpha is a versatile and essential reagent for advancing our understanding of immune regulation, developing new therapies, and investigating the pathogenesis of a wide range of diseases.

You can use recombinant human IL-4 receptor alpha (IL-4Rα) as a standard for quantification or calibration in ELISA assays, provided that the recombinant protein is well-characterized, of high purity, and matches the form of IL-4Rα detected by your assay (e.g., soluble extracellular domain).

Key considerations:

• Recognition by ELISA: Most commercial ELISA kits for human IL-4Rα are designed to detect both endogenous (natural) and recombinant forms of the protein, and their standard curves are typically generated using affinity-purified recombinant IL-4Rα. This means recombinant IL-4Rα is suitable as a standard, as long as it is comparable in structure (e.g., glycosylation, folding) to the native protein in your samples.
• Standard Preparation: The recombinant protein should be reconstituted and diluted according to the manufacturer’s instructions, often in a buffer containing carrier protein (such as BSA) to prevent adsorption and ensure stability.
• Validation: If you are developing your own ELISA or using a non-commercial kit, you should validate the recombinant standard for parallelism with endogenous IL-4Rα in your sample matrix. This ensures that the standard curve accurately reflects the analyte in your samples and that there are no matrix effects or differences in antibody recognition.
• Form of Recombinant Protein: Use the soluble extracellular domain of IL-4Rα, as this is the form typically measured in ELISA assays for soluble receptor quantification. Full-length or membrane-bound forms are not appropriate for soluble receptor quantification.
• Documentation: Ensure the recombinant protein is well-documented (sequence, purity, source, and formulation) to support reproducibility and data interpretation.

Summary Table: Use of Recombinant IL-4Rα as ELISA Standard

Conclusion:
Recombinant human IL-4Rα is widely accepted and used as a standard for ELISA quantification of soluble IL-4Rα, provided it matches the form detected by your assay and is validated for use in your specific application.

Recombinant Human IL-4 Receptor Alpha (IL-4 Rα) has been validated in published research for several key applications, primarily in immunology and inflammation studies:

• Binding Assays and Protein-Protein Interaction Studies: Recombinant IL-4 Rα is widely used to characterize interactions with IL-4, IL-13, and antagonistic antibodies, including epitope mapping and affinity measurements.

• Cellular Functional Assays: It is employed to inhibit IL-4-dependent proliferation of immune cells (e.g., TF-1 cells, T cells, B cells), and to suppress differentiation of naïve CD4+ T cells into Th2 cells, which are central to type 2 immune responses.

• Immunoassays (ELISA, Western Blot): Recombinant IL-4 Rα serves as a standard or capture reagent in ELISA and Western blot protocols for quantifying IL-4, IL-13, or their antagonists, and for pharmacokinetic studies of anti-IL-4Rα antibodies.

• Preclinical Disease Models: It is used in vivo and in vitro to validate therapeutic antibodies targeting IL-4Rα, demonstrating efficacy in models of asthma, atopic dermatitis, and allergic rhinitis by blocking IL-4/IL-13 signaling and reducing inflammation.

• Development of Antagonists and Therapeutics: Recombinant IL-4 Rα is essential for screening and engineering monoclonal antibodies or peptide antagonists that block IL-4/IL-13 signaling, with direct relevance to drug development for allergic and inflammatory diseases.

• Structural and Mechanistic Studies: It is used to study receptor-ligand interactions, receptor complex formation (type I and type II IL-4 receptors), and downstream signaling mechanisms relevant to Th2-biased immunity, alternative macrophage activation, and mucosal immunity.

Summary Table of Validated Applications

These applications are supported by multiple peer-reviewed studies and are foundational for research into type 2 inflammation, allergic diseases, and therapeutic antibody development.

To reconstitute and prepare Recombinant Human IL-4 Receptor Alpha (IL-4 Rα) protein for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Reconstitute the protein in sterile water or PBS. The recommended concentration is typically 0.1–1.0 mg/mL in sterile water or 100 μg/mL in sterile PBS (optionally containing at least 0.1% human or bovine serum albumin for added stability).
• Mix gently by swirling or inverting; do not vortex, as vigorous mixing can denature the protein.
• Allow the solution to sit at room temperature for a few minutes to ensure complete dissolution. If solubility issues persist, incubate at 4°C overnight.
• Filter sterilize the solution if required for your application, using a 0.2 μm filter.

Storage after reconstitution:

• Store at 2–8°C for up to 1 week for short-term use.
• For long-term storage, aliquot and freeze at –20°C to –70°C. Avoid repeated freeze-thaw cycles to maintain protein integrity.

Additional notes for cell culture:

• If using in cell-based assays, dilute the reconstituted stock into your cell culture medium immediately before use.
• If the protein is supplied with carrier protein (e.g., BSA), ensure compatibility with your downstream applications and cell type.
• For Fc-chimera or other fusion constructs, follow the specific reconstitution concentration provided by the manufacturer (e.g., 250 μg/mL in PBS for Fc chimeras).

Summary protocol:

1. Briefly centrifuge vial.
2. Add sterile water or PBS (with or without 0.1% BSA) to achieve 0.1–1.0 mg/mL (or as specified).
3. Gently mix, avoid vortexing.
4. Incubate at room temperature or 4°C if needed.
5. Aliquot and store appropriately.

These steps ensure optimal solubility, stability, and biological activity of recombinant IL-4 Rα for cell culture experiments.