Recombinant Mouse IL-2

Recombinant mouse IL-2 is a valuable tool for immunological research due to its multifaceted roles in immune cell biology and its well-characterized functional properties.

Immune Cell Activation and Proliferation

Recombinant mouse IL-2 serves as a potent lymphoid cell growth factor with primary activity on T cells. It promotes proliferation and differentiation of T cells, NK cells, and B cells, making it essential for studies examining lymphocyte expansion and activation. The cytokine plays a central role in both the activation and maintenance of immune responses and is involved in the elimination of self-reactive T cells.

Regulatory T Cell Development

One of the most significant applications is the study of regulatory T cell (Treg) biology. IL-2 plays a central role in the expansion and maintenance of regulatory T cells, and IL-2 signaling through its receptor complex is critical for thymic Treg differentiation. This makes recombinant mouse IL-2 particularly valuable for research into immune tolerance mechanisms and autoimmune disease models.

Diverse In Vitro Applications

Recombinant mouse IL-2 supports multiple experimental approaches, including:

• T cell activation and expansion: In vitro activation and propagation of mouse T cells using established protocols
• NK cell expansion: In vitro activation and expansion of natural killer cells
• LAK cell generation: Production of lymphokine-activated killer cells for cytotoxicity studies
• Standardization: Use as an IL-2 ELISA standard or Western blot control to ensure reproducibility and precision

Dose-Dependent Functional Flexibility

The functional outcomes of IL-2 treatment are dose-dependent, providing experimental flexibility. While high-dose IL-2 enhances effector T-cell function, low-dose IL-2 unexpectedly induces immune tolerance and promotes Treg development. This dual functionality allows researchers to model different immunological scenarios relevant to cancer immunotherapy, autoimmune disease, and transplantation studies.

Research Reproducibility

Recombinant mouse IL-2 is available in highly purified, premium-grade formulations optimized for research applications, ensuring consistent results across experiments and enabling reliable dose-response studies to determine optimal concentrations for specific applications.

Yes, recombinant Mouse IL-2 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly reconstituted and matched to the assay conditions.

Supporting details:

• Recombinant Mouse IL-2 is commonly used as an ELISA standard for quantifying IL-2 in biological samples. It is specifically formulated and purified for this purpose, ensuring reproducibility and precision in research applications.
• ELISA kits for mouse IL-2 typically include a lyophilized recombinant Mouse IL-2 standard, which is reconstituted and serially diluted to generate a standard curve. This curve is then used to interpolate the concentration of IL-2 in unknown samples.
• The recombinant standard should be reconstituted in the recommended buffer (often phosphate-buffered saline with carrier protein such as BSA) and diluted according to the ELISA protocol to ensure accurate calibration.
• The quantification is valid for both natural and recombinant mouse IL-2, as most ELISA kits are designed to recognize both forms without cross-reactivity to other cytokines.
• For best results, ensure the recombinant IL-2 standard is calibrated against an established reference (such as NIH/NIBSC standard) if absolute quantification is required.

Best practices:

• Always follow the ELISA kit manufacturer’s instructions for reconstitution and dilution of the recombinant standard.
• Prepare a fresh standard curve for each assay to account for any variability in preparation.
• Confirm that the recombinant IL-2 standard is compatible with the antibodies used in your ELISA (capture and detection), as some kits specify particular clones for optimal performance.

Additional notes:

• Recombinant Mouse IL-2 expressed in yeast (e.g., Pichia pastoris) is suitable for ELISA calibration and closely resembles the native protein in terms of folding and post-translational modifications.
• The standard curve should cover the expected range of IL-2 concentrations in your samples, typically from low picogram to nanogram levels, depending on assay sensitivity.

In summary, recombinant Mouse IL-2 is widely accepted and validated as a standard for ELISA quantification, provided it is used according to established protocols and matched to the assay system.

Recombinant Mouse IL-2 has been validated for a range of applications in published research, primarily in immunology and cell biology. The most common validated applications include:

• Cellular bioassays: Used to stimulate and assess the proliferation, activation, or differentiation of T cells, NK cells, and other lymphoid populations in vitro.
• In vivo functional assays: Employed in mouse models to modulate immune responses, such as expanding regulatory T cells (Tregs), enhancing anti-tumor immunity, or promoting transplant tolerance.
• Cell culture supplementation: Added to culture media to support the growth and maintenance of T cells, NK cells, and for differentiation studies.
• ELISA standard: Used as a quantitative standard in immunoassays to measure IL-2 levels.
• Stimulation assays: Applied to stimulate immune cells for downstream functional or phenotypic analysis.

Published research examples:

• Bioassays: Multiple studies have used recombinant mouse IL-2 to stimulate T cells in vitro for proliferation, cytokine production, and cytotoxicity assays.
• In vivo immune modulation: Studies have validated its use in expanding Tregs to promote transplant tolerance and treat autoimmune diseases in mouse models.
• Cancer immunotherapy: Used in preclinical models to enhance anti-tumor immune responses, either alone or in combination with other agents.
• Vaccine adjuvant studies: Incorporated in vaccine formulations to boost cell-mediated and mucosal immunity in mice.

Summary of validated applications in published research:

These applications are well-supported by both product validation data and peer-reviewed research, making recombinant mouse IL-2 a standard tool for immunological studies in mice.

Reconstitution Guidelines

Recombinant Mouse IL-2 is supplied as a lyophilized powder and requires proper reconstitution before use in cell culture applications. The reconstitution process is critical for maintaining protein activity and ensuring reproducible experimental results.

Initial Preparation

Before opening the vial, centrifuge it briefly to bring the lyophilized contents to the bottom. Allow the vial to warm to room temperature without artificial heat (approximately 1 hour) if stored at -80°C. Once warmed, spray the vial with 70% ethanol and place it in a biosafety hood for aseptic reconstitution.

Reconstitution Concentration and Solvent Selection

The recommended reconstitution concentration ranges from 0.1 to 1.0 mg/mL, though most protocols specify 100-200 μg/mL as optimal. The choice of reconstitution solvent depends on your specific product formulation:

• With carrier protein (BSA): Reconstitute in sterile PBS containing at least 0.1% human or bovine serum albumin
• Carrier-free formulations: Reconstitute in sterile deionized water or 100 mM acetic acid
• Alternative option: Sterile 18 MΩ-cm water at concentrations not less than 100 μg/mL

Reconstitution Procedure

Add the appropriate volume of solvent very slowly to minimize splashing and overflow. Gently mix the vial by pipetting the solution down the sides—do not vortex, as this can denature the protein. Allow the solution to sit for approximately 10 minutes to ensure complete reconstitution. If the protein appears as a film at the bottom of the vial, gentle mixing is essential.

Storage and Stability

Short-term Storage

After reconstitution, aliquot the protein into working portions using sterile tubes. Stock solutions can be stored at 2-8°C for up to one week or at -20°C for up to 3 months. For longer-term storage, maintain samples at -80°C.

Critical Storage Considerations

• Use a manual defrost freezer and avoid repeated freeze-thaw cycles
• Store reconstituted protein in sterile conditions
• If using acetic acid for reconstitution, make further dilutions in low endotoxin medium or buffered solutions containing fetal bovine serum or tissue culture-grade BSA

Bioactivity and Quality Control

Recombinant Mouse IL-2 typically has a molecular weight of approximately 17-19 kDa and demonstrates potent bioactivity in cell culture. The protein stimulates proliferation of T cell lines with ED₅₀ values in the range of 0.1-0.4 ng/mL, making it suitable for T cell activation and proliferation assays. Endotoxin levels are typically maintained below 0.1 ng/μg to ensure suitability for sensitive cell culture applications.