Recombinant Human IL-17F

Recombinant Human IL-17F is widely used in research applications to investigate its role as a proinflammatory cytokine, particularly in immune regulation, inflammation, and disease pathogenesis. It is essential for modeling and dissecting the molecular mechanisms underlying autoimmune diseases, host defense, and tissue remodeling.

Key scientific reasons to use Recombinant Human IL-17F in research:

• Modeling Inflammatory Responses: IL-17F stimulates the production of cytokines such as IL-6, IL-8, and G-CSF, and regulates cartilage matrix turnover by increasing matrix release and inhibiting new matrix synthesis. This makes it valuable for studying inflammatory pathways in diseases like rheumatoid arthritis, psoriasis, and inflammatory bowel disease.

• Cellular Signaling Studies: Recombinant IL-17F can be used to activate IL-17RA/RC-dependent intracellular signaling in various cell types, including fibroblasts and endothelial cells, leading to increased expression of cytokines, chemokines, and adhesion molecules. This is critical for understanding leukocyte recruitment and tissue inflammation.

• Synergistic Effects with Other Cytokines: IL-17F, especially when combined with TNF-α, can synergistically regulate gene expression involved in immune cell migration, coagulation, and barrier function. This is important for research into complex immune interactions and xenotransplantation.

• Disease Modeling and Therapeutic Target Validation: Recombinant IL-17F is used in preclinical models to mimic disease states and evaluate the efficacy of therapeutic interventions targeting IL-17 pathways. It helps in validating new drug targets and understanding the molecular basis of disease progression.

• Host Defense Mechanisms: IL-17F plays a role in protective immunity against pathogens by promoting antimicrobial peptide synthesis and supporting neutrophil recruitment. Recombinant protein is used to study these mechanisms in vitro and in vivo.

• Precision Medicine and Biomarker Discovery: Research using IL-17F contributes to identifying biomarkers and therapeutic targets for personalized treatment strategies, especially in autoimmune and inflammatory diseases.

Typical applications include:

• Cell culture stimulation assays to measure cytokine induction.
• Bioassays for immune cell migration and activation.
• Disease modeling in animal and tissue systems.
• Mechanistic studies of gene regulation and signaling pathways.

Using recombinant human IL-17F allows for controlled, reproducible experiments to elucidate its biological functions and therapeutic potential in human health and disease.

Yes, recombinant human IL-17F can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system. This is a common and accepted practice in cytokine quantification.

Key considerations and supporting details:

• Recombinant IL-17F is routinely used as an ELISA standard: Multiple ELISA kits and protocols specify the use of recombinant human IL-17F as the standard for generating calibration curves, enabling quantification of IL-17F in biological samples.
• Assay compatibility: Ensure that the recombinant IL-17F you use matches the specificity of your ELISA antibodies (i.e., the antibodies must recognize the recombinant form equivalently to the native form). Most commercial ELISA kits are validated to detect both natural and recombinant IL-17F.
• Formulation: Recombinant IL-17F is available in different formulations (with or without carrier proteins like BSA). For ELISA standards, formulations containing BSA are often recommended to enhance stability and mimic sample matrix effects.
• No international standard: There is currently no universally accepted international standard for IL-17F calibration, so quantification is typically relative to the recombinant standard provided or specified by the kit or protocol.
• Documentation: Always refer to your ELISA kit’s instructions or published protocols to confirm that the recombinant IL-17F you plan to use is suitable for your specific assay system.

Best practices:

• Reconstitute and dilute the recombinant IL-17F standard according to the ELISA kit or protocol instructions.
• Prepare a standard curve with serial dilutions covering the expected concentration range in your samples.
• Store and handle the recombinant protein as recommended to maintain stability and activity.

Summary:
Recombinant human IL-17F is widely used and appropriate as a standard for ELISA quantification, provided it is compatible with your assay’s antibodies and protocol. Always verify compatibility and follow best practices for preparation and storage.

Recombinant Human IL-17F has been validated in published research for several key applications, primarily in the context of cell-based assays, bioassays, and immunoassays, as well as mechanistic studies of inflammation and immune responses.

Validated Applications in Published Research:

• Cell Culture and Functional Assays:
  IL-17F is widely used to stimulate various human cell types (e.g., airway smooth muscle cells, endothelial cells, adipose stem cells, dermal fibroblasts) to study its effects on cytokine production (such as IL-6, IL-8, G-CSF), chemokine induction, and regulation of matrix turnover. These assays often measure downstream signaling events (e.g., activation of NF-κB, MAPK pathways) and cellular responses like neutrophil adhesion or osteogenic differentiation.

• Bioassays:
  Recombinant IL-17F is used in bioassays to assess its biological activity, such as inducing pro-inflammatory cytokines, modulating immune cell recruitment, and influencing differentiation processes in stem cells. For example, it has been used to demonstrate induction of IL-6 via the TAK1-NFκB pathway in airway smooth muscle cells and to study its role in neutrophil adhesion to endothelial cells.

• ELISA (Enzyme-Linked Immunosorbent Assay):
  IL-17F serves as a standard or capture antigen in ELISA protocols to quantify IL-17F levels in biological samples, such as serum from patients with autoimmune or infectious diseases.

• Disease Mechanism Studies:
  Recombinant IL-17F is used to model and dissect its role in chronic inflammatory diseases (e.g., rheumatoid arthritis, psoriasis, multiple sclerosis), infectious diseases (e.g., COVID-19), and immune deficiencies (e.g., chronic mucocutaneous candidiasis). It is also used to study its effects on endothelial cell activation, angiogenesis, and immune cell recruitment.

• Neutrophil Binding and Adhesion Assays:
  IL-17F has been validated in assays measuring neutrophil adhesion to endothelial cells, demonstrating its role in upregulating adhesion molecules (e.g., ICAM-1) and promoting neutrophil recruitment during inflammation.

Summary Table: Validated Applications

Key Research Contexts:

• Autoimmune and inflammatory diseases: Used to model and understand IL-17F’s role in pathogenesis.
• Infectious diseases: Studied for its elevation and functional effects in conditions like severe COVID-19.
• Tissue remodeling and repair: Investigated for its regulation of cartilage and endothelial cell function.

These applications are supported by multiple peer-reviewed studies and are standard in immunology and cell biology research involving IL-17F.

To reconstitute and prepare Recombinant Human IL-17F protein for cell culture experiments, follow these steps for optimal solubility, stability, and bioactivity:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Reconstitute the protein in a suitable buffer. The most common and recommended buffers are:
  • Sterile 1× PBS (pH 7.4) containing 0.1% endotoxin-free recombinant human serum albumin (HSA).
  • Alternatively, sterile distilled water or aqueous buffer containing 0.1% BSA can be used, especially if albumin is required for stability.
• Recommended concentration for reconstitution is typically 0.1–1.0 mg/mL. For most cell culture applications, 0.2 mg/mL is commonly used.
• Gently mix by swirling or tapping the vial. Do not vortex or pipette vigorously, as this may denature the protein.
• Allow the solution to sit at room temperature for 10 minutes to ensure complete dissolution.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can degrade activity.
• Storage after reconstitution:
  • Short-term (up to 1 week): Store at 2–8°C.
  • Long-term: Store at –20°C to –80°C.
• For cell culture experiments, further dilute the stock solution in your cell culture medium or assay buffer as needed. Ensure the final working concentration matches your experimental design.

Additional notes:

• If the protein is supplied in an acidic buffer (e.g., 4 mM HCl), reconstitute as directed and then dilute into your desired buffer for cell culture.
• Always check the product datasheet or certificate of analysis for specific instructions regarding your batch, as formulation and recommended buffers may vary.
• Avoid repeated freeze-thaw cycles to maintain protein integrity and bioactivity.

Summary of best practices:

• Use sterile PBS + 0.1% HSA/BSA for reconstitution.
• Gentle mixing only.
• Aliquot and store appropriately.
• Dilute into cell culture medium for experiments.

These steps will help ensure the recombinant IL-17F protein remains stable and biologically active for your cell culture assays.