Recombinant Mouse IL-17E

Recombinant Mouse IL-17E (also known as IL-25) is used in research to study its unique roles in immune modulation, inflammation, and tumor biology, making it valuable for investigating Th2-type immune responses, allergic inflammation, and antitumor mechanisms.

Key scientific applications and rationale include:

• Modeling Th2 Immune Responses: IL-17E promotes Th2-biased immune responses, in contrast to other IL-17 family members that drive Th1 or Th17 responses. This makes it essential for dissecting pathways involved in asthma, allergy, and other Th2-mediated diseases.

• Studying Eosinophilic Inflammation: IL-17E induces eosinophilia by stimulating IL-5 production and upregulating IL-4 and IL-13, leading to increased IgA and IgE, mucus production, and epithelial cell hyperplasia. This is relevant for modeling allergic airway inflammation and related pathologies.

• Antitumor Research: Recombinant mouse IL-17E has demonstrated significant antitumor activity in vivo, inhibiting tumor growth in mouse models via multiple administration routes (intraperitoneal, intravenous, subcutaneous). Its efficacy is linked to increased eosinophil infiltration and B cell activation, supporting its use in cancer immunology studies.

• Dissecting Cytokine Signaling: IL-17E binds to the IL-17RB receptor (with IL-17RA as a co-receptor), activating NF-κB and stimulating proinflammatory chemokine (e.g., IL-8) production. This is useful for mechanistic studies of cytokine signaling and immune cell recruitment.

• Comparative Immunology: Mouse IL-17E shares high sequence identity and functional similarity with human IL-17E, allowing translational studies between mouse models and human disease.

• Protein Quality and Consistency: Recombinant production ensures high purity, low endotoxin levels, and batch-to-batch consistency, which are critical for reproducible experimental results.

Summary of research uses:

• Elucidating mechanisms of Th2 immunity and allergic inflammation.
• Investigating the role of eosinophils and B cells in immune responses.
• Exploring antitumor effects and potential immunotherapeutic strategies.
• Studying cytokine signaling pathways relevant to inflammation and immunity.

Using recombinant mouse IL-17E enables precise, controlled studies of these processes, supporting both basic research and preclinical therapeutic development.

Yes, you can use recombinant mouse IL-17E as a standard for quantification or calibration in your ELISA assays, provided it is validated for this purpose in your specific assay system.

Supporting details:

• ELISA kits for mouse IL-17E are routinely calibrated using highly purified recombinant mouse IL-17E as the standard, as stated in multiple kit manuals and protocols. These standards are reconstituted and serially diluted to generate a standard curve for quantification of IL-17E in unknown samples.
• Assay specificity: Most commercial ELISA kits for mouse IL-17E are designed to detect both natural and recombinant forms of the cytokine, and their standard curves are generated using recombinant protein.
• Validation: The use of recombinant mouse IL-17E as a standard is validated by manufacturers for accuracy, linearity, and recovery in various sample matrices (serum, plasma, cell culture supernatant).
• Best practices: Ensure that the recombinant IL-17E you use as a standard is of high purity, correctly folded, and biologically active, as these factors can affect assay performance. It is also important to match the buffer conditions of your standard to those of your samples to minimize matrix effects.

Additional considerations:

• If you are using a commercial ELISA kit, it is recommended to use the recombinant standard provided with the kit or one that matches its specifications, as the assay is validated with that specific standard.
• If you are developing your own ELISA, you can use a recombinant mouse IL-17E standard, but you should validate its performance (e.g., linearity, recovery, parallelism) in your assay system.
• Some suppliers specifically note that their recombinant mouse IL-17E protein is suitable for use as an ELISA standard.

In summary, recombinant mouse IL-17E is the standard material of choice for ELISA quantification of this cytokine, provided it is appropriately validated for your assay conditions.

Recombinant Mouse IL-17E (also known as IL-25) has been validated for several key applications in published research, including bioactivity assays, ELISA, cell culture studies, and in vivo functional studies.

Validated Applications:

• Bioactivity Assays: Recombinant Mouse IL-17E is routinely used to assess its biological activity, such as the induction of cytokine and chemokine expression in target cells, activation of specific signaling pathways, and functional responses in immune cells.

• ELISA Standard: It serves as a standard for quantifying IL-17E levels in biological samples using ELISA, enabling measurement of cytokine concentrations in cell culture supernatants, serum, plasma, and other fluids.

• Cell Culture Studies: The protein is used to stimulate various cell types (e.g., fibroblasts, endothelial cells, immune cells) to study downstream effects such as cytokine production, cell proliferation, and differentiation.

• In Vivo Functional Studies: Recombinant IL-17E has been administered to mice to investigate its role in immune responses, including:

  • Th2 Immune Response Modulation: Promoting Th2-biased immunity, which is critical for defense against helminthic (parasitic worm) infections.
  • Disease Models: Modulating disease severity in models of autoimmune encephalomyelitis (EAE), colitis, and arthritis. Administration of IL-17E has been shown to ameliorate disease in these models by dampening Th1 and Th17 responses.
  • Leukocyte Recruitment: Facilitating recruitment of leukocytes to inflamed tissues in both in vivo (e.g., air pouch model) and in vitro systems.

• Comparative Immunology and Preclinical Therapeutic Evaluation: Used in mouse model studies to evaluate its therapeutic potential and immunomodulatory effects, including comparative studies with other cytokines and therapeutic antibodies.

Supporting Details:

• Assay Platforms: IL-17E is validated for use in single-analyte and multiplex immunoassays, allowing simultaneous measurement with other biomarkers.
• Cell Analysis Methods: It is applied in cell analysis protocols to study gene expression, signaling, and immune cell function.
• Disease Research: IL-17E is investigated for its role in inflammatory diseases, host defense, and tissue repair, as well as its potential as a therapeutic agent for conditions such as IBD, MS, and parasitic infections.

Summary Table of Applications

These applications are widely supported in published research and are foundational for studying the immunological functions and therapeutic potential of recombinant Mouse IL-17E.

To reconstitute and prepare Recombinant Mouse IL-17E (IL-25) protein for cell culture experiments, follow these general best practices based on manufacturer recommendations and scientific protocols:

1. Reconstitution

• Centrifuge the lyophilized protein vial briefly before opening to ensure all powder is at the bottom.
• Reconstitute the protein in sterile, cold distilled water or a mild acidic buffer (such as 10 mM HCl or 4 mM HCl), depending on the product source:
  • For most recombinant proteins (especially those expressed in E. coli), reconstitution in sterile 10 mM HCl at a concentration of ≥100 µg/mL is recommended.
  • For proteins expressed in mammalian cells (e.g., HEK293), reconstitution in sterile distilled water at 0.1–0.5 mg/mL is typical.
• Gently swirl or mix the vial to dissolve the protein. Do not vortex or pipette vigorously to avoid denaturation.

2. Further Dilution

• After reconstitution, dilute the protein to the desired working concentration using cell culture medium (without serum or with low serum, depending on your experiment).
• If the protein is sensitive to aggregation or loss of activity, consider adding a carrier protein (e.g., 0.1% BSA) to the dilution buffer.

3. Aliquoting and Storage

• Aliquot the reconstituted protein into small volumes to minimize freeze-thaw cycles.
• Store aliquots at –20°C to –80°C for long-term storage.
• For short-term use, keep at 2–8°C for up to one week.

4. Handling and Use

• Thaw aliquots on ice and use immediately.
• Avoid repeated freeze-thaw cycles to maintain protein stability and activity.
• For cell culture experiments, add the diluted protein directly to the culture medium.

5. Activity and Concentration

• Typical working concentrations for bioactivity assays range from 0.1–100 ng/mL, depending on the cell type and experimental design.
• Always refer to the product datasheet for specific activity and recommended concentrations.

Example Protocol:

1. Centrifuge the lyophilized vial for 1 min at 10,000 × g.
2. Reconstitute with sterile 10 mM HCl to 100 µg/mL.
3. Gently mix until fully dissolved.
4. Dilute to working concentration (e.g., 10–50 ng/mL) in pre-warmed cell culture medium.
5. Add to cells and incubate as required.

Always check the specific product datasheet for any unique requirements or recommendations.