Recombinant Human IL-17

Recombinant Human IL-17 (particularly IL-17A and IL-17F) is a valuable tool for a wide range of research applications due to its central role in inflammation, immunity, tissue repair, and disease pathogenesis. Here are the key reasons why you should consider using recombinant human IL-17 in your research:

1. Study of Inflammatory and Immune Responses

• IL-17 is a key pro-inflammatory cytokine produced by Th17 cells and other immune cells.
• It drives the expression of numerous chemokines (e.g., CXCL1, CXCL2, CXCL8), cytokines (e.g., IL-1β, IL-6, G-CSF), and antimicrobial peptides, making it essential for modeling inflammatory cascades in vitro and in vivo.
• Useful for investigating the mechanisms of autoimmune diseases (e.g., psoriasis, rheumatoid arthritis, multiple sclerosis) and infectious diseases.

2. Modeling Disease Pathogenesis

• IL-17 is implicated in the development and progression of autoimmune disorders, chronic inflammatory diseases, and certain cancers.
• Recombinant IL-17 allows researchers to mimic disease conditions, study cellular and molecular responses, and test potential therapeutic interventions.

3. Investigation of Tissue Repair and Regeneration

• IL-17 plays a dual role in tissue repair: it can promote acute wound healing and epithelial regeneration, but excessive or prolonged IL-17 signaling may impair healing and contribute to fibrosis.
• Useful for studying wound healing, skin biology, and epithelial barrier function.

4. Drug Screening and Therapeutic Development

• Recombinant IL-17 is used as a standard in assays to screen and validate neutralizing antibodies, small molecule inhibitors, and other therapeutics targeting the IL-17 pathway.
• It is critical for evaluating the efficacy of IL-17 inhibitors (e.g., secukinumab, ixekizumab) and novel biologics in preclinical models.

5. Cell Culture and Functional Assays

• IL-17 can be used to stimulate various cell types (e.g., epithelial cells, fibroblasts, endothelial cells, keratinocytes) to study downstream signaling, gene expression, and functional responses.
• Supports the cultivation and differentiation of musculoskeletal tissues (osteoblasts, cartilage) and skin cells.

6. Mechanistic Studies of IL-17 Signaling

• Enables detailed investigation of IL-17 receptor (IL-17R) signaling pathways, including activation of NF-κB, MAPK, and other downstream effectors.
• Useful for studying post-transcriptional regulation, mRNA stability, and protein translation mechanisms influenced by IL-17.

7. Chemotaxis and Leukocyte Recruitment Assays

• IL-17 induces the production of chemokines that recruit neutrophils and other immune cells, making it ideal for chemotaxis and migration studies.

8. Translational and Preclinical Research

• Recombinant IL-17 is widely used in animal models to study disease mechanisms, evaluate therapeutic strategies, and assess safety and efficacy of new treatments.

Summary:
Recombinant Human IL-17 is indispensable for research on inflammation, immunity, tissue repair, and disease. It enables mechanistic studies, drug screening, and modeling of both protective and pathological processes, making it a versatile and powerful tool for advancing scientific understanding and therapeutic development.

Yes, recombinant human IL-17A is widely used as a standard for quantification or calibration in ELISA assays. This practice is supported by multiple technical sources and is standard in cytokine quantification protocols.

Supporting details:

• Recombinant IL-17A as ELISA Standard: Recombinant human IL-17A is specifically manufactured and validated for use as a standard in ELISA assays. Commercial ELISA kits and protocols routinely use recombinant IL-17A to generate standard curves, enabling accurate quantification of IL-17A in biological samples.
• Parallelism with Native IL-17A: Studies and kit documentation confirm that standard curves generated with recombinant IL-17A are linear and parallel to those obtained with native human IL-17A, allowing reliable quantification of both recombinant and endogenous forms.
• Protocol Considerations: When using recombinant IL-17A as a standard, it is essential to follow the recommended reconstitution and dilution instructions provided by the ELISA kit or protein supplier to ensure accuracy and reproducibility. Standards are typically run in serial dilutions to establish a calibration curve for sample quantification.
• Validation and Application: Recombinant IL-17A standards are validated for use in various sample types, including plasma, serum, and cell culture supernatants, and are compatible with sandwich ELISA formats.

Best Practices:

• Use the recombinant IL-17A standard provided or recommended by your ELISA kit manufacturer for optimal compatibility.
• Prepare the standard curve according to the kit protocol, ensuring proper reconstitution and dilution.
• Confirm that the antibodies used in your assay recognize both recombinant and native IL-17A to ensure accurate quantification.

Summary Table: Recombinant IL-17A as ELISA Standard

In conclusion, recombinant human IL-17A is suitable and routinely used as a standard for ELISA quantification and calibration, provided assay-specific protocols are followed.

Recombinant Human IL-17 (IL-17A) has been validated for a wide range of applications in published research, primarily in bioassays, ELISA development, and functional studies involving immune modulation, inflammation, and disease modeling.

Key validated applications include:

• Bioassays:
  IL-17A is widely used to stimulate various cell types (e.g., HT-29 colon adenocarcinoma cells, human dermal fibroblasts, immune cells) to assess downstream effects such as cytokine and chemokine secretion (e.g., CXCL1/GROα, IL-6), gene expression changes, and cellular responses relevant to inflammation and immunity.

• ELISA Development:
  Recombinant IL-17A serves as a standard for detection and quantification in ELISA assays, enabling measurement of IL-17A levels in biological samples and facilitating the development of diagnostic and research assays.

• Screening of Inhibitory Molecules:
  Used in cell-based assays to screen and validate antibodies or small molecules that block IL-17A signaling, including therapeutic monoclonal antibodies targeting IL-17A (e.g., Secukinumab, Ixekizumab).

• Disease Modeling and Mechanistic Studies:
  IL-17A is employed to model and investigate the pathogenesis of autoimmune and inflammatory diseases such as rheumatoid arthritis, psoriasis, inflammatory bowel disease, asthma, and chronic pain. It is used to induce disease-relevant cellular responses and to study the effects of IL-17A blockade in preclinical models.

• Cell Differentiation and Functional Assays:
  Validated for use in cell culture to study differentiation of immune cells (e.g., Th17 cells, macrophages), phagocytosis, and other functional immune responses.

• Characterization of IL-17A Effects:
  Used to characterize the impact of IL-17A on tissues such as articular cartilage, skin, and vascular endothelium, including induction of inflammatory mediators and tissue remodeling.

• Cancer Research:
  Applied to investigate IL-17A’s role in tumor biology, including its effects on oxidative stress, DNA damage, and immune cell recruitment in cancer cell lines.

Representative published research applications:

• Induction of chemokine secretion (e.g., CXCL1/GROα) in colon cancer cells.
• Promotion of IL-6 production in dermal fibroblasts.
• Cooperative effects with other cytokines (e.g., IL-4) in cancer cell oxidative stress.
• Modeling inflammatory responses in rheumatoid arthritis and IBD.
• Investigation of IL-17A’s role in skin inflammation and wound healing.
• Functional studies in neurological diseases and chronic pain.

Summary Table of Validated Applications

These applications are supported by numerous peer-reviewed studies and are foundational for research into IL-17A’s biological functions, disease relevance, and therapeutic targeting.

To reconstitute and prepare Recombinant Human IL-17 (IL-17A) protein for cell culture experiments, follow these best-practice steps:

1. Briefly centrifuge the vial
This ensures all lyophilized protein is at the bottom before opening.

2. Choose an appropriate reconstitution buffer

• For most cell culture applications, use sterile, endotoxin-free PBS (pH 7.2–7.4) or sterile distilled water.
• If the protein was lyophilized from an acidic solution (e.g., HCl), reconstitute in the same acid (e.g., 4 mM HCl) as recommended by the manufacturer.
• For maximal stability and to prevent adsorption, add a carrier protein such as 0.1–1% BSA or HSA (endotoxin-free).

3. Reconstitution concentration

• Commonly, reconstitute to 0.1–1.0 mg/mL for stock solutions.
• For some protocols, 100 μg/mL is recommended.
• Do not exceed 1 mg/mL to avoid solubility issues.

4. Gently dissolve the protein

• Add the buffer slowly to the vial.
• Gently swirl or tap to mix; do not vortex or shake vigorously to avoid denaturation.
• Allow to sit at room temperature for 10–30 minutes to ensure complete dissolution.

5. Aliquot and storage

• Divide the reconstituted protein into small aliquots to avoid repeated freeze-thaw cycles.
• Store aliquots at –20°C or –70°C for long-term storage; at 2–8°C for up to 1 month if sterile.
• Avoid frost-free freezers and repeated freeze-thawing, which can degrade the protein.

6. Working solution preparation

• For cell culture, dilute the stock solution to the desired working concentration using cell culture medium or buffer containing 0.1% BSA or FBS to minimize adsorption.
• Prepare working solutions fresh before use.

Example protocol for reconstituting lyophilized IL-17A:

1. Briefly centrifuge the vial.2. Add sterile PBS (pH 7.4) with 0.1% BSA to achieve 0.1 mg/mL final concentration.3. Gently swirl to dissolve. Let stand at room temperature for 10–30 minutes.4. Aliquot and store at –20°C or –70°C.5. For experiments, dilute aliquots to working concentrations (e.g., 1–100 ng/mL) in cell culture medium with 0.1% BSA or FBS.

Additional notes:

• Always consult the specific product datasheet for any unique requirements.
• If using for bioassays, confirm activity with a dose-response curve in your system.
• Avoid high concentrations and vigorous mixing to prevent aggregation or loss of activity.

These steps will help ensure protein stability, activity, and reproducibility in your cell culture experiments.