Recombinant Mouse IL-4

Recombinant Mouse IL-4 is widely used in research because it is a key cytokine for modulating immune responses, particularly in studies of B-cell and T-cell biology, macrophage polarization, and allergic inflammation.

IL-4 is essential for:

• Stimulating proliferation of B cells and T cells.
• Inducing immunoglobulin class switching to IgE and IgG1 in B cells, which is critical for humoral immunity and allergy models.
• Driving differentiation of naïve CD4+ T cells into Th2 cells, which are central to adaptive immunity against extracellular pathogens and in allergy research.
• Upregulating MHC class II expression, enhancing antigen presentation.
• Promoting M2 macrophage polarization, which is important for tissue repair, anti-inflammatory responses, and tumor microenvironment studies.
• Supporting mast cell hyperplasia and mucus production, relevant for asthma and allergy models.

Applications include:

• In vitro cell culture: Used to differentiate immune cell subsets (e.g., Th2 cells, dendritic cells from bone marrow precursors), study B cell-T cell interactions, and assess cytokine-driven cell proliferation.
• Functional assays: Investigating cytokine signaling, immune regulation, and cell activation.
• Disease models: IL-4 is used in models of allergy, asthma, autoimmune disease, myocardial infarction (for cardiac repair via M2 macrophages), and cancer immunology.

Using recombinant mouse IL-4 ensures consistent, defined activity and avoids variability inherent in native or serum-derived cytokines, which is crucial for reproducible experimental results.

In summary, recombinant mouse IL-4 is a versatile tool for dissecting immune mechanisms, modeling disease, and manipulating cell fate in vitro and in vivo, making it indispensable for immunology, inflammation, and tissue repair research.

Yes, recombinant mouse IL-4 can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and properly reconstituted.

Recombinant mouse IL-4 is widely used as a standard in ELISA kits for the quantification of IL-4 in biological samples. Commercial ELISA kits specifically calibrate their assays using recombinant mouse IL-4, and studies have shown that dose-response curves for recombinant and natural mouse IL-4 are parallel, indicating that recombinant IL-4 is suitable for accurate quantification. The protein should be reconstituted according to the manufacturer’s instructions, typically in sterile phosphate-buffered saline (PBS) with a carrier protein such as BSA to prevent adsorption and loss of activity.

Key considerations for use as an ELISA standard:

• Purity: Use recombinant IL-4 with >95% purity, confirmed by SDS-PAGE.
• Bioactivity: Ensure the protein is bioactive and structurally similar to native IL-4.
• Reconstitution: Follow recommended protocols for reconstitution, using appropriate diluents and carrier proteins to maintain stability.
• Calibration: Prepare a standard curve with serial dilutions of recombinant IL-4 to enable accurate quantification of unknown samples.
• Compatibility: Confirm that your ELISA kit is validated for recombinant IL-4 standards, as most commercial kits are designed for this purpose.

Limitations:

• Recombinant standards are for research use only and not for diagnostic procedures.
• The standard should be handled and stored according to the supplier’s instructions to preserve activity and accuracy.

In summary, recombinant mouse IL-4 is an established and reliable standard for ELISA quantification, provided you follow best practices for preparation and calibration.

Recombinant Mouse IL-4 has been validated for a broad range of applications in published research, primarily in immunology, cell biology, and disease modeling. Key validated applications include:

• In vitro differentiation of naïve CD4⁺ T cells into Th2 cells.
• Promotion of B cell proliferation and immunoglobulin class switching (IgG1, IgE).
• Functional assays and bioassays to assess cytokine activity, cell proliferation, and immune cell function.
• Bone marrow precursor differentiation into dendritic cells (DCs), often in combination with GM-CSF.
• Investigation of B cell–T cell interactions, including studies with CD40 ligand.
• ELISA standard and Western blot control for quantifying or detecting IL-4 in biological samples.
• Modulation of macrophage polarization and study of M2 macrophage functions in tissue repair, metabolism, and tumor biology.
• Modeling allergic inflammation and asthma, including studies on IgE production and eosinophilic responses.
• Enhancement of antitumor immunity in adoptive cell transfer (ACT) and immune checkpoint blockade (ICB) therapies, including metabolic modulation of CD8⁺ T cells.
• Fibrosis research, particularly in models of tissue remodeling and chronic inflammation.
• Mast cell, basophil, and eosinophil priming and chemotaxis.
• Regulation of dendritic cell and macrophage cytokine production.
• Cell culture supplementation to support immune cell growth and differentiation.

Representative published studies have used recombinant mouse IL-4 for:

• Reprogramming macrophage polarization in cancer and inflammation models.
• Investigating B cell antibody responses and class switching in the absence of T cells.
• Enhancing the efficacy of CAR-T cell therapies and immune checkpoint inhibitors in tumor models.
• Studying the role of IL-4 in tissue repair and metabolic regulation.

Summary Table of Validated Applications

These applications are supported by both product validation data and peer-reviewed publications, confirming the utility of recombinant mouse IL-4 in diverse experimental systems.

To reconstitute and prepare Recombinant Mouse IL-4 protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.

2. Warm the vial to room temperature before opening to minimize condensation.

3. Reconstitution buffer:

   • Use sterile phosphate-buffered saline (PBS) or sterile distilled water as the solvent.
   • For optimal stability and to prevent protein loss, include a carrier protein such as 0.1%–1% Bovine Serum Albumin (BSA) or Human Serum Albumin (HSA) in the buffer.
   • If using directly for cell culture, you may reconstitute in complete cell culture medium containing serum or another acceptable carrier protein.

4. Concentration:

   • Commonly, reconstitute to a stock concentration of 100 μg/mL.
   • Adjust the volume of buffer accordingly based on the amount of lyophilized protein.

5. Dissolving the protein:

   • Add the buffer gently down the side of the vial.
   • Do not vortex; instead, gently pipette up and down or swirl to dissolve.
   • Allow the solution to sit for several minutes at room temperature to ensure complete dissolution.

6. Aliquot and storage:

   • After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles.
   • Store at 2–8°C for up to 1 month or at –20°C to –70°C for up to 3 months under sterile conditions.
   • Avoid repeated freeze-thaw cycles to maintain protein activity.

7. Working solution:

   • Dilute the stock solution to the desired working concentration in cell culture medium immediately before use.
   • Typical working concentrations for cell stimulation are in the ng/mL range (e.g., 0.3–3 ng/mL for bioactivity assays).

Summary Table: Recombinant Mouse IL-4 Reconstitution

Notes:

• Always use sterile technique to avoid contamination.
• If the protein is carrier-free, adding BSA or HSA is highly recommended for stability.
• For specific applications (e.g., ELISA, Western blot), consult the relevant protocol for dilution and buffer requirements.

These steps ensure maximum stability and bioactivity of recombinant mouse IL-4 for cell culture experiments.