Recombinant Human IL-17E

Recombinant Human IL-17E is used in research applications to study and manipulate type 2 immune responses, investigate mechanisms of allergy, asthma, and autoimmune diseases, and explore its antitumor and anti-inflammatory activities.

IL-17E (also known as IL-25) is a cytokine that:

• Induces Th2 cytokines (IL-4, IL-5, IL-9, IL-13), promoting Th2-biased immune responses, which are central to allergic inflammation and defense against helminth infections.
• Suppresses Th1 and Th17 cytokine production, thereby limiting Th17 cell expansion and reducing pathology in autoimmune and inflammatory conditions.
• Amplifies allergic inflammation by increasing serum IgA and IgE, mucus production, epithelial cell hyperplasia, and eosinophilia.
• Exhibits antitumor activity in vivo, as recombinant IL-17E administration in mouse models led to significant tumor growth inhibition, especially when combined with chemotherapy or immunotherapy agents.
• Promotes vascular endothelial cell proliferation and supports blood-brain barrier integrity, relevant for studies in neuroinflammation and tissue remodeling.
• Activates NF-κB and stimulates IL-8 production, making it useful for cell signaling and inflammation studies.
• Is essential for airway remodeling and hyperreactivity, making it a target for respiratory disease research.

Typical research applications include:

• Cell-based bioassays to measure cytokine induction or signaling pathway activation.
• ELISA standards for quantifying IL-17E in biological samples.
• Protein-protein interaction studies to investigate receptor binding and downstream effects.
• In vivo models for allergy, asthma, autoimmune disease, and cancer research.

Using recombinant human IL-17E allows for controlled, reproducible experiments to dissect its biological functions, therapeutic potential, and role in disease pathogenesis.

Yes, recombinant human IL-17E can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this application. Multiple sources confirm that recombinant IL-17E is suitable as an ELISA standard, and commercial ELISA kits are designed to detect both natural and recombinant forms of IL-17E.

Essential context and supporting details:

• Application as ELISA Standard: Recombinant human IL-17E is commonly used as a standard in ELISA assays to generate a calibration curve for quantifying IL-17E concentrations in biological samples. This is standard practice in cytokine quantification, as recombinant proteins provide a defined concentration and purity.

• Validation and Compatibility: It is important to ensure that the recombinant IL-17E you use is compatible with your specific ELISA system. Most commercial ELISA kits specify that their assay will recognize both natural and recombinant human IL-17E, allowing for accurate quantification using recombinant standards. Always refer to the kit insert or technical documentation for confirmation.

• Formulation Considerations: Recombinant IL-17E is often supplied lyophilized with carrier proteins such as BSA to enhance stability. When preparing standards, follow the recommended reconstitution protocol to maintain protein integrity and assay accuracy.

• Quality Control: Recombinant standards should be quality-controlled for purity and biological activity. Commercial sources typically provide lot-specific data and guarantee activity, which is important for reproducibility in quantification.

• Best Practices:

  • Prepare a serial dilution of the recombinant IL-17E to generate a standard curve covering the expected concentration range in your samples.
  • Use the same buffer and matrix for standards and samples to minimize matrix effects.
  • Validate the linearity and sensitivity of your assay using the recombinant standard.

Additional relevant information:

• Species Specificity: Ensure the recombinant IL-17E is human-derived if you are quantifying human samples, as cross-reactivity with other species may affect results.
• Research Use: Most recombinant standards and ELISA kits are intended for research use only and not for diagnostic or therapeutic applications.

Summary Table: Recombinant IL-17E as ELISA Standard

In conclusion, recombinant human IL-17E is appropriate for use as a standard in ELISA quantification, provided it is validated for your assay system and prepared according to recommended protocols.

Recombinant Human IL-17E (also known as IL-25) has been validated for several key applications in published research, primarily in bioassays, ELISA standards, protein-protein interaction studies, and functional assays.

Validated Applications:

• Bioassays:
  IL-17E is widely used in cell-based bioassays to measure its biological activity, such as induction of cytokine secretion (e.g., CXCL1/GROα in HT-29 cells, IL-8 in human PBMCs). These assays assess its ability to modulate immune responses, including Th2 polarization, suppression of Th1/Th17 cytokines, and promotion of angiogenesis.

• ELISA Standard:
  Recombinant IL-17E serves as a standard for quantifying IL-17E levels in human serum and plasma using ELISA, enabling detection and measurement of endogenous protein concentrations.

• Protein-Protein Interaction/Binding Assays:
  It is used in vitro to study interactions with its receptors (IL-17RB, IL-17RA) and downstream signaling pathways, as well as to characterize binding kinetics and functional consequences.

• Functional Assays:
  IL-17E is employed in cell culture and differentiation studies to investigate its effects on immune cell subsets, such as dendritic cells, B cells, and innate lymphoid cells. It has been used to study angiogenesis, blood-brain barrier integrity, and modulation of inflammatory responses.

• In Vivo Antitumor Studies:
  Recombinant IL-17E has demonstrated antitumor efficacy in various human tumor xenograft models (melanoma, breast, lung, colon, pancreatic cancers) in mice, both as a monotherapy and in combination with chemotherapy or immunotherapy agents. These studies involve administration via intraperitoneal, intravenous, or subcutaneous routes and assessment of tumor growth inhibition and immune cell infiltration.

Representative Published Research Applications:

• Induction of cytokine secretion in cell lines (e.g., HT-29, PBMCs).
• Quantification of IL-17E in biological fluids by ELISA.
• Investigation of IL-17E’s role in Th2-biased immune responses, allergic inflammation, airway hyperreactivity, and autoimmune disease models.
• Analysis of angiogenic factor expression and vascular endothelial cell proliferation.
• Assessment of antitumor activity and immune modulation in xenograft models.
• Studies on protein-protein interactions and receptor signaling.

Summary Table of Validated Applications

Additional Notes:

• IL-17E is often used in studies of allergic diseases, autoimmunity, and cancer immunology due to its unique immunomodulatory properties.
• Protocols typically involve recombinant protein reconstitution, dose-response analysis, and endpoint measurements such as cytokine levels, cell proliferation, or histological assessment.

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

To reconstitute and prepare Recombinant Human IL-17E protein for cell culture experiments, follow these best practices based on manufacturer guidelines and scientific protocols:

Reconstitution

1. Centrifuge the vial before opening to ensure the lyophilized powder is at the bottom.
2. Reconstitute the protein in sterile, distilled water or a recommended buffer. For most Recombinant Human IL-17E proteins, the standard reconstitution is:
   • Sterile 4 mM HCl (as recommended by R&D Systems, FUJIFILM Biosciences, and others).
   • Alternatively, sterile distilled water may be used if no specific buffer is indicated (GenScript, ABclonal).
   • Add carrier protein (0.1% human or bovine serum albumin, HSA or BSA) if long-term storage is planned or if the manufacturer recommends it (R&D Systems, Cell Guidance Systems, Assay Biotechnology).
3. Gently mix the solution by pipetting up and down or swirling the vial. Do not vortex to avoid protein denaturation.
4. Reconstitute to a concentration of 100 µg/mL (as per R&D Systems, FUJIFILM Biosciences, Neuromics) or 0.1–0.5 mg/mL (ABclonal, Cell Guidance Systems), depending on the product and intended use.

Preparation for Cell Culture

1. Aliquot the reconstituted protein into small volumes to minimize freeze-thaw cycles.
2. Store aliquots at ≤ –20°C (manual defrost freezer preferred) for long-term storage. Avoid repeated freeze-thaw cycles.
3. For short-term use, store at 2–8°C under sterile conditions for up to 1 month.
4. Dilute the protein in cell culture medium or assay buffer immediately before use. Typical working concentrations for bioassays range from 0.25–1.5 ng/mL, depending on the cell type and experimental design.

Additional Tips

• Always refer to the Certificate of Analysis (COA) or product datasheet for lot-specific instructions.
• For ELISA or binding assays, follow the specific protocol provided by the kit or assay manufacturer.
• If using serum or plasma samples, consider spiking the protein into the appropriate diluent (e.g., Assay Diluent 45HB for serum samples, as per Assay Biotechnology).

By following these steps, you can ensure optimal activity and stability of Recombinant Human IL-17E for your cell culture experiments.