Recombinant Mouse IL-17A/F

Recombinant Mouse IL-17A/F is used in research applications to study immune responses, inflammation, and disease mechanisms because it is a key cytokine involved in both protective immunity and pathological inflammation. The IL-17A/F heterodimer is biologically active and induces chemokine production and neutrophil recruitment, with potency intermediate between IL-17A and IL-17F homodimers.

Key reasons to use recombinant mouse IL-17A/F in research:

• Modeling Inflammatory and Autoimmune Diseases: IL-17A/F is implicated in the pathogenesis of autoimmune and inflammatory diseases such as rheumatoid arthritis, psoriasis, and inflammatory bowel disease. Using recombinant IL-17A/F allows researchers to mimic disease conditions and study cytokine-driven pathology in vitro and in vivo.

• Dissecting Cytokine Signaling Pathways: IL-17A/F activates specific intracellular signaling cascades (e.g., IL-17RA/C-dependent pathways), leading to the expression of cytokines, chemokines, and adhesion molecules in target cells such as fibroblasts and endothelial cells. This is essential for understanding immune cell recruitment and tissue inflammation.

• Studying Immune Cell Function: IL-17A/F modulates the activity of various immune cells, including macrophages and CD8+ T cells, influencing their cytokine profiles, activation states, and effector functions. For example, IL-17A/F can promote the shift of macrophages toward a pro-inflammatory phenotype and enhance CD8+ T cell cytotoxicity.

• Host Defense and Infection Models: IL-17A/F is critical for mucosal and epidermal defense against bacterial and fungal infections, promoting neutrophil influx and antimicrobial peptide production. Recombinant cytokine is used to probe these mechanisms in infection models.

• Therapeutic Target Validation: Recombinant IL-17A/F is used to test the efficacy of neutralizing antibodies or small molecules targeting the IL-17 pathway, supporting preclinical drug development and mechanistic studies.

• Standardization and Reproducibility: Recombinant proteins provide a consistent, defined reagent for controlled experiments, reducing variability compared to native or crude cytokine preparations.

Typical applications include:

• Cell culture stimulation assays to induce cytokine/chemokine production.
• In vivo administration to model inflammation or infection.
• Functional assays to study immune cell activation, migration, or cytotoxicity.
• Screening and validation of IL-17-targeted therapeutics.

In summary, recombinant mouse IL-17A/F is a versatile tool for dissecting the roles of IL-17 family cytokines in immunity, inflammation, and disease, enabling both basic research and translational studies.

Yes, recombinant mouse IL-17A/F can be used as a standard for quantification or calibration in ELISA assays, provided the assay is designed to detect the IL-17A/F heterodimer and the standard is appropriately validated for your specific kit and protocol.

Supporting details:

• Assay Compatibility: Multiple ELISA kits specifically designed for mouse IL-17A/F heterodimer use recombinant mouse IL-17A/F as the calibration standard. These kits demonstrate that the recombinant protein produces standard curves that are parallel to those generated with natural IL-17A/F, indicating suitability for quantification.
• Calibration and Quantification: The standard curve in these assays is generated using serial dilutions of recombinant mouse IL-17A/F, allowing for accurate quantification of IL-17A/F in unknown samples. The concentration of IL-17A/F in samples is determined by comparing their signal to this standard curve.
• Specificity: These ELISAs are validated to recognize both natural and recombinant mouse IL-17A/F heterodimer, with no significant cross-reactivity or interference from related cytokines.
• Best Practices: Ensure that the recombinant standard you use matches the form of IL-17A/F detected by your assay (i.e., heterodimer, not IL-17A or IL-17F homodimers), and that it is prepared and stored according to the manufacturer’s recommendations for consistency and accuracy.

Limitations and Considerations:

• Research Use Only: Most commercial ELISA kits and recombinant standards are for research use only and not for diagnostic applications.
• Validation: If using a recombinant standard from a different source than the kit manufacturer, it is advisable to validate that the standard curve generated is parallel to that of the kit’s recommended standard, ensuring accurate quantification.
• Matrix Effects: Recovery and precision data indicate that recombinant IL-17A/F performs reliably in various sample types (serum, plasma, cell culture supernatant), but matrix effects should always be considered and, if necessary, validated for your specific application.

Summary Table: Use of Recombinant Mouse IL-17A/F as ELISA Standard

Conclusion:
You can use recombinant mouse IL-17A/F as a standard for quantification or calibration in ELISA assays, provided the assay is validated for this purpose and the standard is handled according to best laboratory practices.

Recombinant Mouse IL-17A/F has been validated for several key applications in published research, primarily in bioassays, in vivo studies, and as a stimulus in cell culture models to study immune and inflammatory responses.

Validated Applications in Published Research:

• Bioassays:
  Recombinant Mouse IL-17A/F is widely used in bioassays to assess its biological activity, such as the induction of chemokine production, neutrophil recruitment, and cytokine secretion (e.g., IL-6 induction in NIH/3T3 cells). These assays are fundamental for characterizing the functional properties of IL-17A/F in vitro.

• In Vivo Studies:
  The protein has been used in mouse models to study its role in inflammation, mucosal immunity, airway neutrophilia, and disease models such as arthritis, multiple sclerosis, psoriasis, and transplant rejection. For example, it has been administered to mice to induce or modulate inflammatory responses and to evaluate therapeutic interventions targeting the IL-17 pathway.

• Cell Culture Stimulation:
  IL-17A/F is applied to primary cells or cell lines (e.g., fibroblasts, macrophages, stromal cells) to stimulate signaling pathways, gene expression (such as antimicrobial peptides), and cytokine/chemokine production. This is essential for dissecting downstream effects and mechanisms of IL-17 signaling.

• ELISA Standard:
  Recombinant Mouse IL-17A (and by extension, IL-17A/F) is validated as a standard in sandwich ELISA assays for quantifying IL-17 levels in biological samples.

Representative Research Contexts:

• Autoimmune and Inflammatory Disease Models:
  Used in preclinical models of rheumatoid arthritis, inflammatory bowel disease, and airway inflammation to study disease mechanisms and evaluate anti-IL-17 therapeutics.

• Mucosal Immunity and Host Defense:
  Applied to investigate the induction of antimicrobial peptides and the recruitment of immune cells in response to bacterial infections.

• Mechanistic Studies:
  Utilized to activate specific signaling pathways (e.g., IL-17RA/RC-dependent signaling) in vitro and in vivo, enabling the study of downstream molecular and cellular events.

Summary Table:

Key Notes:

• The IL-17A/F heterodimer is biologically active and exhibits intermediate potency between IL-17A and IL-17F homodimers, making it particularly relevant for comparative studies of IL-17 family cytokines.
• Applications are supported by both direct experimental use and as a reference standard for assay development and validation.

If you require protocols or more detailed application notes for a specific experimental context, please specify the intended use.

To reconstitute and prepare Recombinant Mouse IL-17A/F protein for cell culture experiments, dissolve the lyophilized protein in sterile 4 mM HCl containing at least 0.1% human or bovine serum albumin (BSA or HSA) to a final concentration of 100 μg/mL. This carrier protein is essential to stabilize the cytokine and prevent adsorption to tube walls, especially at low working concentrations.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile 4 mM HCl containing 0.1% BSA or HSA to the vial to achieve the desired stock concentration (typically 100 μg/mL).
• Gently pipet up and down and wash down the sides of the vial to ensure complete solubilization.
• Allow several minutes for complete reconstitution; avoid vigorous vortexing to prevent protein denaturation.
• Aliquot the stock solution to minimize freeze-thaw cycles. Store aliquots at −80°C for long-term storage.
• For cell culture experiments, dilute the stock solution further in cell culture medium or assay buffer containing 0.1% carrier protein to prevent loss of activity due to adsorption.

Additional notes:

• Always use sterile technique to avoid contamination.
• Avoid repeated freeze-thaw cycles, as these can degrade the protein.
• If the protein is provided in a different buffer or formulation, consult the specific product datasheet for recommended reconstitution conditions.
• For short-term storage (days), keep at 2–8°C; for long-term, store at −80°C.
• If using for ELISA or other assays, follow the specific dilution and buffer recommendations provided in the assay protocol.

This protocol ensures optimal solubility, stability, and biological activity of recombinant Mouse IL-17A/F for cell culture applications.