Recombinant Mouse IL-15

Recombinant Mouse IL-15 is widely used in research applications due to its potent ability to stimulate and expand key immune cell populations, particularly CD8+ T cells and natural killer (NK) cells, making it essential for studies in immunology, cancer immunotherapy, and infectious disease models.

IL-15 is a cytokine that plays a central role in both innate and adaptive immunity by:

• Promoting the activation, proliferation, and survival of NK cells, NKT cells, and CD8+ T cells.
• Supporting the maintenance and homeostasis of memory T cells, which is crucial for long-term immune protection and vaccine studies.
• Inducing or enhancing the differentiation and activation of multiple T cell subsets, including Th17, Treg, and CD8+ memory cells.

Key research applications include:

• Cell culture and differentiation studies: Recombinant Mouse IL-15 is optimized for in vitro expansion and activation of NK cells and T cells, including differentiation of NK cells from progenitors and expansion of naïve CD8+ T cells.
• Functional assays: Used to assess cytotoxicity, cytokine production, and immune cell function in response to various stimuli.
• Cancer immunotherapy models: Systemic administration of recombinant IL-15 in mice has been shown to promote tumor regression, decrease metastasis, and increase survival, often by enhancing cytotoxicity of CD8+ T cells and NK cells. IL-15 can also be used in combination with other immunotherapies for synergistic anti-tumor effects.
• Infectious disease and vaccine studies: IL-15 is critical for the development of protective immune responses against microbial invaders and parasites, and for the migration of effector CD8+ T cells to infection sites.
• Inflammation and autoimmunity research: IL-15 is implicated in the pathogenesis of several inflammatory disorders, making it a target for studies on immune regulation and therapeutic intervention.

Advantages of recombinant protein use:

• Consistency and reproducibility: Recombinant Mouse IL-15 provides a defined, consistent cytokine source, minimizing variability in experimental outcomes.
• Species specificity: Mouse IL-15 is ideal for murine models, ensuring compatibility with mouse immune cells and accurate physiological responses.

In summary, Recombinant Mouse IL-15 is a versatile tool for immunological research, enabling precise manipulation of immune cell populations and facilitating studies in cancer, infectious disease, inflammation, and immune regulation.

Yes, recombinant mouse IL-15 can be used as a standard for quantification or calibration in ELISA assays, provided it is suitable for your specific assay format and detection system.

Recombinant mouse IL-15 (such as those from PeproTech, Boster Bio, or BioLegend) is typically produced as a purified protein and is commonly used to generate standard curves in ELISA kits for mouse IL-15. These standards allow for the accurate quantification of endogenous IL-15 in samples by comparing the signal from your samples to the known concentrations of the recombinant protein.

However, there are a few important considerations:

• Purity and Activity: Ensure the recombinant IL-15 is of high purity and has been validated for use in immunoassays.
• Compatibility: Confirm that the recombinant protein is compatible with the antibodies and detection system used in your ELISA kit. Some kits may specify which recombinant standards are recommended.
• Bioassay vs. ELISA: Recombinant proteins intended for ELISA standards may not be suitable for bioassays unless specifically tested and validated for biological activity.

In summary, recombinant mouse IL-15 is appropriate for use as a standard in ELISA assays for quantification, but always check the manufacturer’s recommendations and validate its performance in your specific assay conditions.

Recombinant Mouse IL-15 has been validated for a wide range of applications in published research, primarily in immunology and oncology. The most common validated applications include:

• Cell culture and functional assays: Used to stimulate, expand, and differentiate various immune cell populations, especially NK cells, NKT cells, and CD8+ T cells in vitro.
• Bioassays: Frequently employed in proliferation and cytotoxicity assays, such as the CTLL-2 cell proliferation assay to measure biological activity.
• In vivo studies: Used in mouse models to study immune cell development, memory T cell maintenance, antitumor immunity, and immune responses to infection or vaccination.
• Antitumor and immunotherapy research: Validated for enhancing antitumor immunity, both as a monotherapy and in combination with other agents (e.g., checkpoint inhibitors, nanovaccines), and for improving CAR-NK cell persistence and function.
• Differentiation studies: Supports in vitro differentiation of NK cells from progenitors and influences the development and function of T cell subsets, including Th17, Treg, and memory CD8+ T cells.
• Survival and anti-apoptotic signaling: Demonstrated to deliver survival signals to NK, T, and B cells, supporting their maintenance in culture and in vivo.
• Migration and trafficking studies: Used to investigate the migration of effector CD8+ T cells, such as their recruitment to lung airways after infection.
• Vaccine adjuvant research: Explored as an adjuvant to enhance antigen-specific T cell responses in vaccine models.

Supporting details from published research:

• In vitro: IL-15 is used for the activation and expansion of NK and NKT cells, differentiation of NK cells from CD34+ progenitors, and expansion/activation of naive CD8+ T cells and cytotoxic T lymphocytes (CTLs).
• In vivo: Studies have validated IL-15 for promoting memory CD8+ T cell survival, enhancing antitumor immunity, and supporting durable immune responses in tumor and infection models.
• Bioassays: The ED50 for recombinant mouse IL-15 is commonly determined using CTLL-2 cell proliferation assays, confirming its biological activity.
• Immunotherapy: IL-15 has been used to improve the persistence and effectiveness of CAR-NK cells and as a component of biomimetic nanovaccines for cancer therapy.

Summary of validated applications:

• Cell culture (immune cell expansion, differentiation)
• Functional and proliferation bioassays
• In vivo immunology and oncology models
• Antitumor and immunotherapy studies
• Vaccine adjuvant research
• Migration/trafficking studies

These applications are well-supported by both product validation data and peer-reviewed publications, demonstrating the versatility of recombinant mouse IL-15 in basic and translational immunology research.

To reconstitute and prepare Recombinant Mouse IL-15 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer to a concentration of 100 μg/mL. Use either sterile water or sterile PBS, and if possible, include at least 0.1% carrier protein (such as human or bovine serum albumin) to stabilize the cytokine and minimize adsorption to plasticware.

Step-by-step protocol:

• Centrifuge the vial briefly (e.g., 3000 rpm for 5 min) before opening to ensure all powder is at the bottom.
• Add sterile buffer:
  • For carrier-containing formulations: Use sterile PBS with ≥0.1% BSA.
  • For carrier-free formulations: Use sterile water or PBS; addition of 0.1% BSA is recommended for extended storage.
• Reconstitute to 100 μg/mL:
  • Calculate the volume needed based on the mass of protein in the vial.
  • Gently mix by pipetting; do not vortex or shake vigorously, as this may reduce biological activity.
• Incubate at room temperature for at least 20 minutes to ensure complete dissolution.
• Aliquot and store:
  • Short-term: 2–8 °C for up to 1 month.
  • Long-term: –20 °C to –70 °C, avoiding repeated freeze-thaw cycles.
  • For carrier-free protein, add 0.1% BSA before freezing to improve stability.

Preparation for cell culture:

• Dilute the stock solution in cell culture medium to the desired working concentration immediately before use.
• Typical working concentrations for bioassays range from 0.1–10 ng/mL, depending on cell type and experimental design.
• Always use sterile technique to avoid contamination.

Additional notes:

• Avoid vigorous mixing or vortexing to preserve protein activity.
• If the protein is supplied with a Certificate of Analysis (CoA), follow the specific reconstitution instructions provided.
• For sensitive applications, confirm protein activity using a proliferation assay (e.g., CTLL-2 cells).

This protocol ensures optimal solubility, stability, and biological activity of recombinant mouse IL-15 for cell culture experiments.