Recombinant Rat IL-17AF

Recombinant Rat IL-17AF is a heterodimeric cytokine composed of IL-17A and IL-17F subunits, and its use in research is crucial for studying inflammatory responses, immune regulation, and disease mechanisms in rat models.

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

• Modeling Inflammatory Pathways: IL-17AF signals through the IL-17RA/IL-17RC receptor complex, regulating the expression of chemokines and cytokines that drive neutrophil recruitment and tissue inflammation. This is essential for investigating the molecular basis of diseases such as autoimmune disorders, asthma, and pulmonary fibrosis.

• Studying Disease Mechanisms: IL-17AF is implicated in the pathogenesis of chronic inflammatory diseases, including psoriasis, rheumatoid arthritis, and interstitial lung disease. Using recombinant rat IL-17AF allows for controlled in vitro and in vivo experiments to dissect its role in disease progression and resolution.

• Therapeutic Target Validation: Since IL-17A and IL-17F are major targets for anti-inflammatory therapies, recombinant IL-17AF is valuable for preclinical testing of inhibitors, antibodies, or small molecules that modulate IL-17 signaling.

• Functional Assays: Recombinant rat IL-17AF can be used to stimulate cells (e.g., fibroblasts, epithelial cells, immune cells) to assess downstream effects such as IL-6 secretion, chemokine production, and activation of signaling pathways like NF-κB.

• Species-Specific Research: Rat models are widely used in immunology and disease studies. Recombinant rat IL-17AF ensures species compatibility, avoiding cross-reactivity issues that may arise with human or mouse cytokines.

Typical applications include:

• In vitro cell stimulation assays to measure cytokine and chemokine induction.
• In vivo administration to model inflammatory or autoimmune diseases.
• Mechanistic studies of IL-17 signaling in rat tissues and primary cells.
• Validation of therapeutic agents targeting IL-17A/F pathways.

Best practices:

• Use recombinant rat IL-17AF at concentrations validated for your assay type.
• Include appropriate controls (e.g., vehicle, single subunit cytokines) to distinguish heterodimer-specific effects.
• Confirm receptor expression (IL-17RA/IL-17RC) in your cell or tissue model for optimal responsiveness.

In summary, recombinant rat IL-17AF is a critical tool for dissecting the biology of IL-17-mediated inflammation and for translational research aimed at developing new therapies for immune-mediated diseases.

You can use recombinant rat IL-17AF (IL-17A/F heterodimer) as a standard for quantification or calibration in ELISA assays, but only if your ELISA is specifically validated to detect the IL-17A/F heterodimer, not just IL-17A or IL-17F homodimers.

Key considerations:

• ELISA specificity: Most commercial rat IL-17A ELISA kits are designed to detect the IL-17A homodimer, not the IL-17A/F heterodimer. These kits typically use antibodies that are highly specific for IL-17A and show minimal cross-reactivity with IL-17A/F or IL-17F. For example, one kit reports only ~2.2% cross-reactivity with recombinant mouse IL-17A/F in a rat IL-17A assay. This means using IL-17AF as a standard in an IL-17A-specific ELISA would result in significant underestimation of concentrations, as the assay would not efficiently detect the heterodimer.

• Assay validation: If you have an ELISA specifically designed or validated for the IL-17A/F heterodimer (sometimes called IL-17AF), then recombinant rat IL-17AF is appropriate as a standard. Some kits or custom assays are available for this purpose and will specify that they detect the heterodimer.

• Biological relevance: IL-17A, IL-17F, and IL-17A/F are distinct molecular species with different biological activities and immunoreactivities. The standard used must match the analyte you intend to quantify.

Best practices:

• Match standard to analyte: Use recombinant rat IL-17A as a standard for IL-17A-specific ELISAs, recombinant rat IL-17F for IL-17F-specific ELISAs, and recombinant rat IL-17AF for assays validated to detect the heterodimer.
• Check kit documentation: Always consult your ELISA kit’s documentation for information on cross-reactivity and recommended standards. Do not substitute standards unless the assay is validated for the alternative form.

Summary Table:

If your ELISA is not specifically validated for the IL-17A/F heterodimer, do not use recombinant rat IL-17AF as a standard for quantification or calibration, as this will compromise the accuracy of your results.

Recombinant Rat IL-17AF has been validated in published research for several key applications, primarily related to its role as an effector cytokine in immune and inflammatory responses.

Validated Applications in Published Research:

• In Vivo Functional Studies:
  Recombinant rat IL-17AF has been used in animal models to study its biological effects. For example, chronic infusion of IL-17 (including IL-17AF) in pregnant rats was shown to promote hypertension, activate cytolytic natural killer cells, and induce vascular dysfunction, demonstrating its utility in in vivo disease modeling and mechanistic studies of inflammation and immune modulation.

• Cell-Based Bioactivity Assays:
  IL-17AF has been validated for inducing cytokine secretion (such as IL-6) in fibroblast cell lines (e.g., NIH-3T3 mouse embryonic fibroblasts), confirming its bioactivity and suitability for in vitro assays measuring downstream signaling and cytokine induction.

• Immunological and Inflammatory Pathway Analysis:
  IL-17AF is widely used to investigate its signaling through the IL-17RA/IL-17RC receptor complex, leading to activation of NF-κB and MAPK pathways, and subsequent transcription of cytokines, chemokines, and antimicrobial peptides. These studies often use recombinant IL-17AF to dissect its role in neutrophil recruitment, chemokine production, and tissue inflammation in both in vitro and in vivo systems.

• Disease Model Research:
  Recombinant rat IL-17AF has been applied in models of inflammatory diseases such as arthritis, asthma, and autoimmune disorders to study its contribution to disease severity and immune cell recruitment.

Additional Context:

• Cross-Species Activity:
  Recombinant rat IL-17AF is active on mouse cells, making it suitable for use in murine models as well as rat systems.

• Mechanistic Studies:
  It is used to elucidate the distinct and overlapping functions of IL-17A, IL-17F, and the IL-17AF heterodimer, particularly in studies comparing their relative potencies in inducing chemokines and neutrophilia.

• Potential for Screening:
  Recombinant IL-17AF is also employed in screening assays for inhibitors or modulators of the IL-17 pathway, relevant for therapeutic development.

Summary Table: Applications of Recombinant Rat IL-17AF

These applications are supported by both product validation data and peer-reviewed research, establishing recombinant rat IL-17AF as a versatile tool for immunological and inflammatory research.

To reconstitute and prepare Recombinant Rat IL-17AF protein for cell culture experiments, dissolve the lyophilized protein in sterile water at a concentration between 0.1 mg/mL and 1 mg/mL. After reconstitution, further dilute the protein in your desired cell culture medium or buffer for experimental use.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all powder is at the bottom.
• Add sterile water to the vial to achieve a final concentration of 0.1–1 mg/mL (commonly 0.1 mg/mL is used for most applications).
• Gently pipette up and down and wash down the sides of the vial to ensure complete dissolution of the protein.
• Avoid vigorous mixing to prevent protein denaturation or foaming.
• Let the solution stand at room temperature for 15–30 minutes with gentle agitation to ensure full solubilization.
• If required for stability, add a carrier protein such as 0.1% BSA or HSA to the solution, especially for long-term storage or repeated freeze-thaw cycles.
• Aliquot the solution to avoid repeated freeze-thaw cycles, and store at 4°C for short-term (2–7 days) or below –18°C for long-term.
• For cell culture experiments, dilute the reconstituted stock to the desired working concentration in your cell culture medium. The effective concentration for bioactivity (e.g., induction of IL-6 in NIH-3T3 cells) is typically in the range of 3–23 ng/mL, depending on assay conditions.

Additional notes:

• Always use sterile technique to prevent contamination.
• If the protein is shipped with a carrier (e.g., BSA), reconstitute in PBS as recommended by the manufacturer; otherwise, sterile water is suitable.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.
• For long-term storage, aliquot and freeze at –20°C to –80°C, optionally with 5–50% glycerol for added stability.

Summary Table:

This protocol ensures optimal recovery and activity of recombinant Rat IL-17AF for cell culture experiments.