Recombinant Mouse IL-17

Recombinant Mouse IL-17 (typically IL-17A) is used in research to model and dissect the biological functions of this cytokine in mouse systems, particularly its roles in inflammation, immune response, and disease pathogenesis. It enables controlled, reproducible studies of IL-17-mediated signaling and cellular responses in vitro and in vivo.

Key reasons to use recombinant mouse IL-17 in research applications:

• Modeling Inflammatory Pathways: IL-17A is a potent proinflammatory cytokine involved in the recruitment of neutrophils, induction of chemokines, and amplification of immune responses, making it essential for studying mechanisms of inflammation, autoimmunity, and host defense.
• Disease Mechanism Studies: Recombinant IL-17A is widely used to mimic or exacerbate disease states in mouse models, such as ischemic stroke, multiple sclerosis, rheumatoid arthritis, and inflammatory bowel disease, allowing researchers to investigate the cytokine’s pathological roles and test therapeutic interventions.
• Cellular and Molecular Assays: It is used in cell culture to stimulate immune or non-immune cells, enabling analysis of downstream signaling, gene expression, cytokine production, and functional responses such as apoptosis or proliferation.
• Standardization and Reproducibility: Recombinant proteins provide a consistent, defined source of cytokine, ensuring reproducibility across experiments and between laboratories, which is critical for quantitative assays (e.g., ELISA standards, bioassays, Western blot controls).
• Stem Cell and Tissue Repair Research: IL-17A influences the differentiation and function of hematopoietic and mesenchymal stem cells, making recombinant IL-17 valuable for studies on tissue repair, regeneration, and stem cell biology.
• Therapeutic Target Validation: By adding recombinant IL-17A to experimental systems, researchers can validate the effects of IL-17 blockade or neutralization, supporting the development of IL-17-targeted therapies for autoimmune and inflammatory diseases.

Experimental examples:

• In a mouse ischemic stroke model, recombinant mouse IL-17A was shown to aggravate neuronal injury and apoptosis in a dose-dependent manner, demonstrating its utility in modeling neuroinflammatory damage and testing neuroprotective strategies.
• In vitro, IL-17A increases proliferation and colony formation of bone marrow-derived mesenchymal stem cells, supporting its use in stem cell and regenerative medicine research.

Summary of applications:

• In vivo disease modeling (e.g., stroke, arthritis, infection)
• In vitro cell stimulation and signaling studies
• ELISA and assay standardization
• Stem cell differentiation and proliferation assays
• Validation of anti-IL-17 therapeutics

Using recombinant mouse IL-17 allows precise control over experimental conditions and is essential for dissecting the cytokine’s complex roles in health and disease.

Yes, recombinant mouse IL-17 can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose in your specific assay system. Multiple commercial ELISA kits and protocols use recombinant mouse IL-17 as the standard to generate calibration curves for quantifying IL-17 in biological samples.

Key supporting details:

• Assay Validation: Commercial mouse IL-17 ELISA kits are designed to quantitate both recombinant and natural mouse IL-17. These kits typically use recombinant mouse IL-17 as the standard, and validation data show that the standard curves generated with recombinant protein are parallel to those obtained with natural IL-17, indicating comparable quantification.
• Application in ELISA: Recombinant mouse IL-17 is explicitly recommended as a standard for sandwich ELISA assays by multiple sources. For example, BioLegend and R&D Systems provide recombinant mouse IL-17A specifically for use as an ELISA standard.
• Performance: The use of recombinant IL-17 as a standard allows for accurate and reproducible quantification of IL-17 in various sample types, including serum, plasma, and cell culture supernatants.
• Best Practices: Ensure that the recombinant IL-17 standard is of high purity, correctly folded, and matched to the isoform detected by your assay antibodies. Always prepare and handle the standard according to the ELISA kit or protocol instructions to maintain accuracy and reproducibility.

Additional considerations:

• If you are developing a custom ELISA, confirm that your capture and detection antibodies recognize both recombinant and native forms equivalently.
• For absolute quantification, ensure the recombinant standard is well-characterized and quantified (e.g., by amino acid analysis or absorbance at 280 nm).

In summary, recombinant mouse IL-17 is widely accepted and validated as a standard for ELISA quantification, provided it is compatible with your assay system and antibodies.

Recombinant Mouse IL-17 (typically IL-17A) has been validated for a range of applications in published research, including bioassays, in vivo studies, ELISA standards, and cell culture experiments.

Key validated applications include:

• Bioassays:
  Recombinant mouse IL-17A is widely used in cell-based bioassays to study its biological activity, such as induction of cytokine production (e.g., IL-6 in NIH/3T3 cells), nitric oxide synthase expression, and signaling pathway activation in various cell types.

• In Vivo Studies:
  It has been administered to mice to model and investigate its role in disease processes, including inflammation, autoimmune diseases (e.g., arthritis, multiple sclerosis, hepatitis), neuroinflammation, and tissue injury.

• ELISA Standard:
  Recombinant mouse IL-17A is validated as a standard for quantitative measurement of IL-17A in mouse samples using sandwich ELISA assays.

• Cell Culture Experiments:
  Used to stimulate primary cells or cell lines (e.g., fibroblasts, macrophages, T cells) to assess downstream effects such as cytokine secretion, gene expression, and cell signaling responses.

• Functional Studies:
  Applied in studies to dissect mechanisms of immune regulation, T cell polarization (e.g., Th17 differentiation), and to evaluate the effects of IL-17A on other immune cell subsets.

Representative published research applications:

• Cardiomyocyte studies: Investigating IL-17A-mediated signaling and cell death in cardiac cells.
• Neuroinflammation and neurodegeneration: Modeling the role of IL-17A in multiple sclerosis and Alzheimer’s disease in mice.
• Autoimmune and inflammatory disease models: Studying arthritis, hepatitis, and airway inflammation in vivo.
• ELISA and cytokine quantification: Serving as a standard for quantifying IL-17A in biological samples.
• Cell signaling and cytokine induction: Inducing IL-6 and other cytokines in fibroblasts and stromal cells.

Summary Table of Validated Applications

These applications are supported by multiple peer-reviewed studies and product validation data, confirming the utility of recombinant mouse IL-17A in both basic and translational immunology research.

To reconstitute and prepare Recombinant Mouse IL-17 protein for cell culture experiments, dissolve the lyophilized protein in sterile, distilled water to achieve a concentration between 0.1–1.0 mg/mL. For optimal stability and biological activity, further dilute the stock solution in a buffered solution containing a carrier protein such as heat-inactivated fetal calf serum (FCS) or tissue culture grade bovine serum albumin (BSA).

Step-by-step protocol:

• Add sterile distilled water directly to the lyophilized IL-17 protein to reach your desired stock concentration (typically 0.1–1.0 mg/mL).
• Gently mix by pipetting up and down or by slow vortexing. Avoid vigorous agitation to prevent protein denaturation.
• Allow the solution to sit at room temperature for a few minutes to ensure complete dissolution.
• For working concentrations, dilute the stock solution in cell culture medium or a buffered solution containing a carrier protein (e.g., 0.1% BSA or heat-inactivated FCS) to minimize adsorption and loss of activity.
• Filter sterilize the final working solution if necessary, using a low protein-binding filter (e.g., 0.2 μm PVDF).
• Use immediately or aliquot and store at 2–8°C for up to one month, or at –20°C to –70°C for longer-term storage. Avoid repeated freeze-thaw cycles to preserve protein integrity.

Additional notes:

• Always use low endotoxin reagents and buffers to prevent unwanted immune activation in cell culture.
• If the formulation contains additives (e.g., acetonitrile, TFA, or BSA), follow the manufacturer’s reconstitution instructions, which may specify using acidic buffers (e.g., 4 mM HCl) and carrier proteins for stability.
• Confirm the biological activity of the reconstituted protein using a relevant bioassay (e.g., induction of IL-6 in NIH/3T3 cells).

This protocol ensures the recombinant mouse IL-17 protein is properly solubilized, stabilized, and ready for use in cell culture experiments.