Recombinant Human IL-31

Recombinant Human IL-31 is used in research to study its roles in inflammation, immune signaling, and disease mechanisms, particularly in skin and airway disorders. It enables controlled, reproducible experiments on IL-31’s biological effects and signaling pathways.

Key reasons to use recombinant human IL-31 in research applications:

• Modeling Inflammatory and Allergic Diseases: IL-31 is a pro-inflammatory cytokine implicated in conditions such as atopic dermatitis, allergic contact dermatitis, and asthma. Using recombinant IL-31 allows researchers to mimic disease-relevant cytokine environments in vitro and in vivo, facilitating the study of disease mechanisms and therapeutic interventions.

• Dissecting Signal Transduction Pathways: Recombinant IL-31 activates key signaling cascades, including the JAK/STAT, PI3K/AKT, and MAP kinase pathways, through its receptor complex (IL-31RA/OSMRβ). This makes it valuable for investigating downstream molecular events and gene expression changes in target cells.

• Studying Chemokine and Cytokine Induction: IL-31 stimulates the production of chemokines (e.g., CCL2/MCP-1) and cytokines (e.g., IL-6, IL-8, EGF, VEGF) in epithelial cells and immune cells, which are critical for recruiting inflammatory cells and mediating tissue responses. This is essential for understanding immune cell migration and tissue inflammation.

• Therapeutic Target Validation: Recombinant IL-31 is used to screen and validate potential inhibitors, monoclonal antibodies, or small molecules targeting IL-31 or its receptor, supporting drug discovery for inflammatory and pruritic diseases.

• Functional and Structural Studies: It enables biochemical and structural analyses of IL-31-receptor interactions, aiding in the design of receptor antagonists or modified cytokines with altered activity.

• Neuro-Immune Research: IL-31 directly activates sensory neurons involved in itch (pruritus), making it a tool for studying neurogenic inflammation and sensory neuron biology.

Summary of applications:

Using recombinant human IL-31 ensures experimental reproducibility, defined cytokine concentrations, and species specificity, which are critical for mechanistic studies and translational research.

Yes, recombinant human IL-31 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated and matched to your assay system. This is a common and accepted practice in cytokine ELISA protocols.

Supporting details:

• ELISA kits for human IL-31 routinely use recombinant human IL-31 as the standard for generating the calibration curve, against which unknown sample concentrations are determined. These standards are typically supplied as lyophilized recombinant protein, which is reconstituted and serially diluted to create the standard curve.
• Both natural and recombinant IL-31 are recognized by validated ELISA kits, and the quantification is based on the standard curve generated from the recombinant standard.
• Carrier protein considerations: Recombinant IL-31 is often supplied with a carrier protein (such as BSA) to enhance stability, especially for use as an ELISA standard. Carrier-free versions are available if BSA or other additives could interfere with your assay.
• Validation: It is important to ensure that the recombinant IL-31 you use as a standard is compatible with your specific ELISA antibodies and detection system. Most commercial ELISA kits are optimized for their supplied standard, but if you are developing your own assay or substituting a standard, you should verify that the recombinant protein is recognized equivalently to the native protein in your samples.
• Standard curve preparation: Always prepare a fresh standard curve for each assay, using serial dilutions of the recombinant IL-31 standard as described in your ELISA protocol.

Best practices:

• Use the same buffer and diluent for your standard as for your samples to minimize matrix effects.
• Confirm the concentration and purity of your recombinant IL-31 standard.
• Store and handle the recombinant protein according to manufacturer or protocol recommendations to maintain stability and activity.

In summary: Recombinant human IL-31 is widely used and appropriate as a standard for ELISA quantification, as long as it is validated for your assay system and handled according to best practices.

Recombinant Human IL-31 has been validated for several key applications in published research, primarily in studies of cytokine signaling, immune cell activation, and disease modeling related to inflammation and pruritus.

Validated Applications:

• Cell Culture and Bioassays:
  Recombinant Human IL-31 is widely used to stimulate various cell types in vitro, including human airway smooth muscle cells, intestinal epithelial cells, and bronchial epithelial cell lines (e.g., BEAS-2B, A549). It has been shown to activate signaling pathways such as Jak/STAT, PI3K/AKT, and MAP kinase, and to induce secretion of chemokines like CCL2/MCP-1.

• Signal Transduction Studies:
  IL-31 is validated for use in assays measuring STAT3 phosphorylation and downstream signaling events, particularly in epithelial and immune cells expressing IL-31RA and OSMRβ. These assays are critical for dissecting the molecular mechanisms of IL-31-mediated inflammation.

• Binding and Receptor Interaction Assays:
  Recombinant IL-31 has been used in ELISA and flow cytometry to confirm binding to its receptor IL-31RA and OSMRβ, both in recombinant and cell-surface contexts. Surface plasmon resonance (SPR) has also been employed to quantify binding affinities.

• Immunogenicity and Vaccine Development:
  IL-31-derived peptides and recombinant protein have been used to develop and validate vaccine candidates targeting chronic pruritus and dermatological disorders. These studies assess antibody titers, avidity, and the ability of vaccine-induced antibodies to inhibit IL-31 signaling in vitro.

• Disease Modeling:
  Recombinant IL-31 has been used in animal models (e.g., transgenic mice) to induce phenotypes such as pruritus, alopecia, and skin lesions, supporting its role in atopic dermatitis and other inflammatory skin diseases. Humanized mouse models expressing IL-31 and its receptors have been validated for functional studies in atopic dermatitis.

• Structural and Therapeutic Studies:
  Purified recombinant IL-31 is used for structural analysis and as a tool in the development of therapeutic agents targeting IL-31-related pathways.

Summary Table of Validated Applications

Additional Notes:

• IL-31 is most commonly used in studies of inflammatory skin diseases (e.g., atopic dermatitis), pruritus, and allergic responses.
• It is validated for use with whole cells, recombinant receptor proteins, and in vivo models.
• The protein is typically expressed in E. coli or mammalian systems and is available in forms suitable for both biochemical and cell-based assays.

If you require protocols or specific assay details for any of these applications, please specify the experimental context.

To reconstitute and prepare Recombinant Human IL-31 protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial:
Briefly spin the vial to ensure all lyophilized protein is at the bottom before opening.

2. Reconstitution:

• Use sterile PBS (phosphate-buffered saline) as the solvent.
• For most applications, reconstitute to a concentration between 0.1–0.5 mg/mL (100–500 μg/mL). Some protocols recommend reconstitution at 10–200 μg/mL depending on your experimental needs.
• If the protein is supplied with a carrier (e.g., BSA), reconstitute in PBS containing at least 0.1% BSA to stabilize the protein and prevent adsorption to surfaces.

Example protocol:

Add sterile PBS (with 0.1% BSA if not already present) to the vial to achieve your desired concentration (e.g., 100 μg/mL). Gently pipette the solution down the sides of the vial to dissolve the protein. Do not vortex.

3. Aliquoting and Storage:

• Divide the reconstituted protein into small aliquots to avoid repeated freeze-thaw cycles, which can degrade the protein.
• Store aliquots at ≤ –20°C for long-term storage.
• For short-term use (2–7 days), store at 4–8°C.

4. Working Dilutions:

• Prepare further dilutions in cell culture medium or buffered solution containing carrier protein (e.g., PBS with 0.1% BSA) immediately before use.
• Avoid repeated freeze-thaw cycles by thawing only the amount needed for each experiment.

5. Handling Tips:

• Gently mix by pipetting; do not vortex, as this can denature the protein.
• If the solution is not clear, allow it to sit at room temperature for a few minutes and gently invert to mix.

Summary Table:

Note: Always consult the specific product datasheet for any unique instructions, as formulations and recommended concentrations may vary between suppliers.