Recombinant Human IL-17 R

Using Recombinant Human IL-17 Receptor (IL-17 R) in research applications is essential for studying the IL-17 signaling pathway, which is central to inflammation, autoimmunity, and host defense. This recombinant protein enables precise investigation of ligand-receptor interactions, screening of inhibitors, and functional assays relevant to disease mechanisms.

Key reasons to use Recombinant Human IL-17 R include:

• Mechanistic Studies: IL-17 R is the primary receptor for IL-17 cytokines (notably IL-17A and IL-17F), mediating downstream signaling that induces pro-inflammatory cytokines (e.g., IL-6, TNFα), chemokines (e.g., CXCL1, CCL20), and matrix metalloproteinases. Using the recombinant receptor allows you to dissect these pathways in vitro and in cell-based assays.

• Drug Discovery and Screening: Recombinant IL-17 R is widely used to screen and characterize small molecules, antibodies, or peptides that block IL-17/IL-17R interactions—a validated therapeutic strategy for autoimmune diseases such as psoriasis and rheumatoid arthritis. It serves as a target or capture reagent in binding assays, ELISA, and high-throughput screening platforms.

• Functional Assays: The recombinant receptor can be used to neutralize IL-17 activity in cell culture, enabling assessment of IL-17-dependent responses such as cytokine secretion, cell migration, and proliferation. For example, it can inhibit IL-17-induced IL-6 secretion in fibroblast assays, providing a quantitative readout of pathway blockade.

• Biophysical and Structural Studies: Recombinant IL-17 R is suitable for binding affinity measurements (e.g., SPR, BLI), crystallography, and mapping of ligand-receptor interfaces, which are critical for rational drug design and understanding receptor activation mechanisms.

• Modeling Disease Pathogenesis: Since IL-17 R is highly expressed on immune cells, fibroblasts, and keratinocytes, recombinant forms are used to model disease-relevant interactions and to generate IL-17R-deficient or overexpressing cell lines for comparative studies.

• Quality Control and Standardization: Recombinant IL-17 R provides a consistent, defined reagent for assay calibration, antibody validation, and as a standard in quantification assays, ensuring reproducibility across experiments.

In summary, Recombinant Human IL-17 R is a versatile tool for elucidating IL-17 biology, validating therapeutic targets, and developing new interventions for inflammatory and autoimmune diseases.

Recombinant Human IL-17 R (IL-17RA/IL-17R) can be used as a standard for quantification or calibration in ELISA assays, provided the assay is specifically designed to measure IL-17 R and not IL-17A. The suitability depends on the target analyte and the assay configuration.

Essential Context and Supporting Details

• ELISA Standard Use: Recombinant proteins are commonly used as standards in ELISA assays to generate calibration curves for quantification. For IL-17A quantification, recombinant human IL-17A is used as the standard. For IL-17R quantification, recombinant human IL-17R (IL-17RA) can be used as the standard, but only in assays designed to detect IL-17R.

• Assay Target Specificity:

  • If your ELISA is designed to detect IL-17A (the cytokine), you must use recombinant IL-17A as the standard.
  • If your ELISA is designed to detect IL-17R (the receptor), recombinant IL-17R is appropriate as the standard.

• Protein Formulation: Recombinant IL-17R is available in formulations suitable for use as ELISA standards. For example, the biotinylated recombinant human IL-17RA/IL-17R Fc chimera is recommended for use as an ELISA standard.

• Calibration Curve: The standard protein should be serially diluted to generate a calibration curve, allowing quantification of unknown samples by interpolation.

Additional Relevant Information

• Assay Validation: Ensure the recombinant IL-17R standard is validated for your specific ELISA protocol. The standard should match the epitope recognized by the capture and detection antibodies in your assay.

• Matrix Effects: When quantifying IL-17R in biological samples, confirm that recovery and linearity are acceptable in your sample matrix (e.g., serum, plasma, cell culture supernatant).

• Not Interchangeable: Do not use recombinant IL-17R as a standard for IL-17A quantification, or vice versa, as these are distinct proteins with different biological roles and immunoreactivity.

Summary Table: Appropriate Standard for ELISA Target

In conclusion: Use recombinant human IL-17R as a standard only for ELISAs designed to quantify IL-17R, not IL-17A. Always verify compatibility with your assay's antibody pair and protocol.

Recombinant Human IL-17 R (IL-17RA) has been validated for several key applications in published research, primarily in the context of immunology, inflammation, and autoimmune disease studies.

Validated Applications:

• Functional ELISA: Used to measure binding interactions with IL-17A and IL-17F, and to quantify ligand-receptor affinity.
• Cell-based Assays: Employed to assess inhibition of IL-17-induced cytokine secretion (e.g., IL-6 production in NIH-3T3 fibroblasts).
• Western Blot (SDS-PAGE): Used for protein characterization, purity assessment, and molecular weight determination.
• Flow Cytometry: Applied for detection and quantification of IL-17RA expression on cell surfaces, and for validation of knockdown or overexpression models.
• Neutralization Assays: Used to evaluate the ability of recombinant IL-17RA to block IL-17A/F signaling and downstream inflammatory responses.
• Binding Activity Studies: Validated for ligand-receptor binding, including studies of receptor-ligand specificity and affinity.
• In Vivo and In Vitro Functional Studies: Utilized in models of autoimmune diseases (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis) to investigate the role of IL-17RA in disease progression and immune cell recruitment.

Supporting Details:

• ELISA Standards: Recombinant IL-17RA is commonly used as a standard for quantifying IL-17A/F in biological samples.
• Cell Culture: Carrier-free forms are recommended for cell or tissue culture applications, including studies of cytokine signaling and immune cell activation.
• Therapeutic Target Validation: Recombinant IL-17RA has been used to validate drug candidates and biologics targeting the IL-17 pathway in preclinical and translational research.
• Disease Models: Research has demonstrated its utility in models of arthritis, psoriasis, and other inflammatory conditions, where IL-17RA blockade or modulation affects disease outcomes.

Summary Table of Validated Applications

Additional Notes:

• Recombinant IL-17RA is a critical reagent for dissecting IL-17 signaling mechanisms, validating therapeutic targets, and studying immune responses in both basic and translational research.
• Its use spans from molecular assays to complex disease models, reflecting its central role in IL-17-mediated pathophysiology.

If you require protocol details or specific experimental setups for any of these applications, please specify the context or research focus.

To reconstitute and prepare Recombinant Human IL-17 R (IL-17RA) protein for cell culture experiments, use sterile phosphate-buffered saline (PBS) as the diluent and aim for a final concentration of 100–250 μg/mL depending on your experimental requirements.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Add sterile PBS (pH 7.2–7.4) to the vial to achieve the desired concentration (commonly 100–250 μg/mL).
• Gently mix by swirling or tapping the vial. Avoid vigorous pipetting or vortexing to prevent protein denaturation.
• Allow the protein to dissolve at room temperature for 10–15 minutes. Inspect visually to confirm complete dissolution.
• If recommended by your protocol, add a carrier protein such as 0.1–1% endotoxin-free human serum albumin (HSA) or bovine serum albumin (BSA) to improve stability, especially for low concentrations or repeated freeze-thaw cycles.
• Filter sterilize the solution if required for cell culture, using a 0.2 μm syringe filter.

Storage and handling:

• After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles.
• Store aliquots at 2–8°C for up to 1 week or at –20°C to –70°C for longer-term storage.
• Avoid storing diluted protein for extended periods; use freshly prepared solutions for optimal activity.

Additional notes:

• Always consult the specific Certificate of Analysis (CoA) or product datasheet for your recombinant protein batch for any unique instructions.
• For functional assays, typical working concentrations range from 5–100 ng/mL depending on the cell type and experimental design.
• Ensure all buffers and reagents are endotoxin-free to prevent unwanted cell activation.

This protocol is suitable for preparing recombinant IL-17 R for cell stimulation, blocking assays, or binding studies in cell culture.