Recombinant Rat IL-17F

Recombinant Rat IL-17F is used in research to study immune regulation, inflammation, and disease mechanisms, particularly those involving Th17 cells and chronic inflammatory conditions. It is a critical tool for dissecting the biological functions of IL-17F in rat models and for evaluating therapeutic strategies targeting the IL-17 pathway.

Key scientific reasons to use recombinant rat IL-17F in your research include:

• Modeling Inflammatory and Autoimmune Diseases: IL-17F is a pro-inflammatory cytokine produced mainly by Th17 cells. It plays a significant role in the pathogenesis of autoimmune diseases, chronic inflammation, and infection models. Using recombinant IL-17F allows you to mimic or modulate these disease processes in vitro or in vivo, facilitating the study of disease mechanisms and potential interventions.

• Functional Assays and Mechanistic Studies: Recombinant rat IL-17F can be used to stimulate cells (e.g., fibroblasts, epithelial cells, immune cells) to assess downstream signaling, cytokine production (such as IL-6), and gene expression changes. For example, it has been shown to induce IL-6 production in mouse fibroblast cells in a dose-dependent manner, making it valuable for mechanistic studies of cytokine signaling.

• Comparative Studies with IL-17A and Heterodimers: IL-17F shares significant homology with IL-17A and can form IL-17A/F heterodimers, each with distinct biological activities. Using recombinant IL-17F enables direct comparison of the effects of IL-17A, IL-17F, and their heterodimers on target cells, which is important for understanding their individual and combined roles in immune responses.

• Therapeutic Target Validation: Dysregulation of IL-17F is implicated in diseases such as psoriasis, psoriatic arthritis, and certain cancers. Recombinant IL-17F is used to validate therapeutic targets, screen inhibitors (such as neutralizing antibodies), and evaluate the efficacy of dual IL-17A/IL-17F blockade strategies.

• Standardization and Quantification: Recombinant IL-17F serves as a standard in ELISA and other immunoassays for quantifying endogenous IL-17F levels in biological samples, ensuring assay accuracy and reproducibility.

• Species-Specific Research: Using recombinant rat IL-17F ensures species compatibility in rat models, which is essential for translational relevance and accurate interpretation of immunological data.

In summary, recombinant rat IL-17F is a versatile reagent for investigating the role of IL-17F in immune regulation, disease pathogenesis, and therapeutic intervention in rat-based research systems. Its use is fundamental for both basic and translational studies targeting the IL-17 signaling axis.

Yes, recombinant rat IL-17F protein can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated and matched to the assay system. Recombinant cytokines are commonly used as standards in quantitative sandwich ELISA protocols for cytokine measurement, including IL-17F.

Essential context and supporting details:

• ELISA Standard Curve: Quantitative ELISA assays require a standard curve generated using known concentrations of the target analyte. Recombinant rat IL-17F is suitable for this purpose, as it provides a defined concentration and purity, allowing accurate interpolation of sample concentrations.
• Assay Compatibility: It is critical that the recombinant IL-17F standard is compatible with the antibodies used in your ELISA kit. Most commercial rat IL-17F ELISA kits specify that their standard is recombinant rat IL-17F, and the assay is validated to detect both natural and recombinant forms.
• Validation: Ensure the recombinant protein is bioactive and structurally similar to native IL-17F. Some suppliers confirm bioactivity by functional assays, such as induction of IL-6 in responsive cell lines. The standard should be reconstituted and diluted according to the kit protocol to match the matrix and conditions of your samples.
• Best Practices:
  • Prepare the standard curve in duplicate or triplicate for accuracy.
  • Use the same buffer/matrix for standards and samples to minimize matrix effects.
  • Do not mix reagents from different kits or lots, as antibody specificity and standard protein formulation may vary.
  • Confirm that your ELISA kit detects recombinant IL-17F with similar sensitivity as native protein.

Additional relevant information:

• Carrier Proteins: Some recombinant standards are supplied with carrier proteins (e.g., BSA) to stabilize the protein. Choose the formulation recommended for ELISA standard use.
• Bioassay vs. ELISA Standard: Recombinant proteins validated for ELISA standard use may not be suitable for bioassays unless specifically tested for biological activity.
• Documentation: Refer to your ELISA kit’s protocol for recommended standard preparation and dilution ranges. Most kits provide a lyophilized recombinant IL-17F standard for this purpose.

In summary, recombinant rat IL-17F is widely accepted as a standard for ELISA quantification, provided it is validated for use with your specific assay system and prepared according to best practices.

Recombinant Rat IL-17F has been validated for several key applications in published research, primarily in functional assays, binding activity, and SDS-PAGE.

Validated Applications:

• Functional Assays:
  Recombinant rat IL-17F is widely used to assess its biological activity, such as its ability to induce cytokine production (e.g., IL-6) in cell-based assays. For example, it has been shown to induce IL-6 production in fetal mouse skin fibroblast cells in a dose-dependent manner. This is a standard method to confirm the protein's bioactivity.

• Binding Activity:
  The protein has been validated for binding activity, which is essential for studying receptor interactions and downstream signaling pathways.

• SDS-PAGE:
  SDS-PAGE is routinely used to confirm the purity and molecular weight of recombinant rat IL-17F, with reported purities exceeding 97%.

Additional Context and Research Uses:

• In Vivo Studies:
  Recombinant rat IL-17F has been used in animal models to study its role in immune responses, such as the induction of neutrophilia in the lungs and exacerbation of antigen-induced pulmonary inflammation. These studies help elucidate IL-17F's function in disease models, including inflammatory and autoimmune conditions.

• Mechanistic Studies:
  IL-17F is a critical effector cytokine involved in host defense, neutrophil recruitment, and epithelial barrier maintenance. Recombinant forms are used to dissect these mechanisms in vitro and in vivo.

• Disease Models:
  Research applications include investigating IL-17F's role in autoimmune diseases, infection, cancer, and tissue integrity. Recombinant protein is used to stimulate cells or tissues to model disease processes and immune responses.

Summary Table of Validated Applications

Note:
While recombinant rat IL-17F is primarily validated for the above applications, it is also used as a standard in ELISA assays and for mechanistic studies in immunology and inflammation research, though direct validation for ELISA is more commonly reported for IL-17A or IL-17A/F heterodimers. Always consult specific published protocols for details on experimental conditions and validation.

To reconstitute and prepare Recombinant Rat IL-17F protein for cell culture experiments, follow these best-practice steps:

1. Equilibrate: Allow the lyophilized protein vial and your chosen reconstitution buffer (commonly sterile PBS or sterile distilled water) to reach room temperature before opening to prevent condensation.

2. Centrifuge: Briefly centrifuge the vial to collect all lyophilized material at the bottom.

3. Reconstitution:

   • Add sterile buffer (commonly PBS, pH 7.2–7.4, or sterile distilled water) to achieve your desired concentration. A typical starting concentration is 100–200 μg/mL.
   • For example, add enough buffer to reach 100 μg/mL if using for cell culture. If your protocol or datasheet specifies a different concentration, follow that guidance.
   • Gently pipette up and down or swirl (do not vortex) to dissolve the protein completely. Let the solution sit at room temperature for 15–30 minutes with gentle agitation to ensure full dissolution.

4. Carrier Protein (Optional but Recommended for Storage):

   • For long-term storage or to prevent adsorption to tube walls, add a carrier protein such as 0.1% BSA or HSA.

5. Aliquot and Storage:

   • Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
   • Store at 4°C for short-term use (up to 1 week); for long-term storage, keep at –20°C or below.
   • Avoid repeated freeze-thaw cycles, as this can reduce protein activity.

6. Preparation for Cell Culture:

   • Before use, dilute the reconstituted stock to the desired working concentration using sterile cell culture medium or PBS. Typical working concentrations for functional assays (e.g., cytokine stimulation) are in the ng/mL range (e.g., 1–100 ng/mL), depending on the assay and cell type.
   • Filter sterilize the final working solution if sterility is critical and the buffer composition allows.

Summary Table: Key Steps for Recombinant Rat IL-17F Reconstitution

Additional Notes:

• Always consult the specific product datasheet for any unique instructions or buffer requirements, as formulations may vary.
• If the protein is supplied with additives (e.g., BSA), reconstitute as directed by the manufacturer.
• For functional assays, verify bioactivity using a relevant cell-based assay (e.g., IL-6 induction in NIH-3T3 cells).

These steps will ensure optimal solubility, stability, and biological activity of recombinant rat IL-17F for cell culture applications.