Recombinant Mouse IL-20

Recombinant Mouse IL-20 is used in research applications to study its role as a cytokine in regulating inflammation, immune responses, and tissue homeostasis, particularly in models of skin disease, autoimmune disorders, and hematopoiesis.

IL-20 is a member of the IL-20 receptor cytokine family, which includes IL-19, IL-22, and IL-24. These cytokines are important for mediating communication between immune cells and epithelial cells, influencing both innate and adaptive immunity. Specifically, IL-20 regulates the proliferation and differentiation of keratinocytes during inflammation, making it highly relevant for research into skin diseases such as psoriasis, where it is critical for both induction and maintenance of disease pathology. Continuous administration of recombinant IL-20 in animal models can induce psoriasis-like lesions, while blockade of IL-20 can resolve these lesions, demonstrating its direct involvement in disease mechanisms.

Beyond dermatological research, IL-20 also promotes the expansion of multipotential hematopoietic progenitor cells, making it useful for studies on hematopoiesis and immune cell development. In autoimmune disease models, IL-20 and related cytokines have been shown to regulate T cell responses, modulate cytokine production, and affect the balance between pro-inflammatory and anti-inflammatory pathways.

Key research applications for recombinant mouse IL-20 include:

• Modeling and dissecting mechanisms of chronic inflammation and autoimmune diseases (e.g., psoriasis, rheumatoid arthritis, inflammatory bowel disease).
• Studying epithelial-immune cell crosstalk and its impact on tissue homeostasis and host defense.
• Investigating hematopoietic progenitor cell biology and immune cell differentiation.
• Testing therapeutic strategies that target IL-20 signaling pathways for disease intervention.

When using recombinant mouse IL-20, it is important to select preparations with high purity and validated biological activity, as determined by cell proliferation assays and endotoxin testing. This ensures reproducibility and reliability in experimental outcomes.

In summary, recombinant mouse IL-20 is a valuable tool for elucidating cytokine-driven mechanisms in inflammation, immunity, and tissue pathology, with particular relevance to skin and autoimmune disease research.

Yes, recombinant Mouse IL-20 can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately known. This is a common practice in quantitative ELISA protocols for cytokines such as IL-20.

Key considerations and supporting details:

• Standard Curve Requirement: Quantitative ELISA assays require a standard curve generated from known concentrations of the target analyte, typically using recombinant protein standards. This allows for the interpolation of sample concentrations based on their optical density (OD) readings relative to the standard curve.

• Recombinant Protein as Standard: Most commercial Mouse IL-20 ELISA kits use recombinant Mouse IL-20 as the standard for calibration. The recombinant protein should be well-characterized, with a defined concentration and high purity, to ensure accurate quantification.

• Matrix Effects: The standard is usually diluted in a buffer or diluent provided or recommended by the ELISA kit to match the sample matrix as closely as possible, minimizing matrix effects that could impact quantification accuracy.

• Validation: It is important to confirm that the recombinant Mouse IL-20 you intend to use is compatible with your specific ELISA assay, especially regarding its source, folding, and post-translational modifications, as these can affect antibody recognition. Ideally, use a recombinant standard that matches the one validated for your ELISA kit.

• Documentation: Ensure that the recombinant Mouse IL-20 is supplied with a certificate of analysis or datasheet specifying its concentration, purity, and biological activity, as these parameters are critical for reliable standard curve generation.

Protocol best practices:

• Prepare a serial dilution of the recombinant Mouse IL-20 in the recommended diluent to generate a standard curve covering the expected range of your samples.
• Run standards in duplicate or triplicate to ensure reproducibility.
• Always include a fresh standard curve with each assay to account for potential day-to-day variability.

Summary Table: Use of Recombinant Mouse IL-20 as ELISA Standard

In conclusion, recombinant Mouse IL-20 is suitable as a standard for ELISA quantification, provided it meets the above criteria and is used according to best practices for standard curve preparation and assay validation.

Recombinant Mouse IL-20 has been validated in published research primarily for applications related to cell-based functional assays, in vitro cell proliferation and differentiation studies, and in vivo disease modeling, especially in the context of inflammation and skin biology.

Key validated applications include:

• Cell Proliferation Assays: Recombinant Mouse IL-20 has been used to stimulate keratinocyte proliferation and differentiation in vitro, which is relevant for studying inflammatory skin diseases such as psoriasis. The biological activity is commonly validated by measuring cell proliferation, with reported ED₅₀ values in the range of 0.5–2.5 ng/mL.

• In Vivo Disease Models: IL-20 has been used in mouse models to investigate its role in the induction and maintenance of psoriasis and other inflammatory conditions. Overexpression or administration of recombinant IL-20 in transgenic mice has been shown to promote keratinocyte hyperproliferation and prevent terminal differentiation, mimicking features of human psoriasis.

• Hematopoietic Cell Expansion: IL-20 has been shown to cause expansion of multipotential hematopoietic progenitor cells, indicating its use in studies of hematopoiesis and immune cell development.

• Inflammation and Cytokine Signaling Studies: Recombinant Mouse IL-20 is used to dissect signaling pathways in keratinocytes and other epithelial cells, particularly those involving the IL-20 receptor complexes.

• ELISA Standard and Positive Control: The protein is frequently used as a standard or positive control in ELISA and other immunoassays to quantify IL-20 or to validate antibody specificity.

• Cell/Tissue Culture Supplement: It is used as a supplement in cell or tissue culture to study cytokine-mediated effects on various cell types, especially epithelial and immune cells.

Supporting details from published research:

• IL-20’s role in skin inflammation and psoriasis has been validated in mouse models, where it regulates keratinocyte proliferation and differentiation.
• Functional validation is typically performed using cell proliferation assays and in vivo models of skin disease.
• The protein’s activity is confirmed by its ability to induce expected biological responses in target cells, such as keratinocytes.

Additional notes:

• While some sources mention related cytokines (e.g., IL-22) in liver disease models, the direct use of recombinant mouse IL-20 in these contexts is less well documented in the provided search results.
• Recombinant Mouse IL-20 is not typically used for diagnostic or therapeutic purposes in humans; its applications are restricted to preclinical research.

Summary of validated applications:

• Cell-based functional assays (proliferation, differentiation)
• In vivo disease modeling (especially skin inflammation/psoriasis)
• Hematopoietic progenitor cell expansion studies
• ELISA and immunoassay standard/positive control
• Cell/tissue culture supplement for cytokine signaling research

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

To reconstitute and prepare Recombinant Mouse IL-20 protein for cell culture experiments, follow these best practices based on manufacturer guidelines and scientific protocols:

Reconstitution

1. Solvent: Reconstitute the lyophilized protein in sterile, endotoxin-free distilled water (ddH₂O) or sterile H₂O.
2. Concentration: Prepare a stock solution at a concentration of not less than 100 μg/mL (0.1 mg/mL), or within the range of 0.1–0.5 mg/mL as recommended by several suppliers.
3. Procedure:
   • Gently add the appropriate volume of sterile water to the vial.
   • Allow the vial to sit for a few minutes to let the powder hydrate.
   • Gently mix by swirling or pipetting up and down. Do not vortex, as this may denature the protein.
   • Incubate the solution at room temperature for at least 20 minutes to ensure complete dissolution.

Storage of Stock Solution

• Short-term: Store the reconstituted stock at 2–8°C for up to 1 week (some sources allow up to 1 month).
• Long-term: Aliquot and store at –20°C to –70°C.
• Avoid repeated freeze-thaw cycles to maintain protein activity.

Preparation for Cell Culture

1. Dilution:
   • Dilute the stock solution in low endotoxin cell culture medium or a buffered solution (e.g., PBS with carrier protein).
   • For sensitive assays, consider adding a carrier protein (e.g., 0.1% BSA, 10% FBS, or 5% HSA) to stabilize the cytokine.
2. Working Concentration:
   • The typical working concentration for bioactivity assays is 0.5–2.5 ng/mL (ED₅₀), but optimize based on your experimental system.
3. Sterility:
   • Ensure all steps are performed under aseptic conditions to avoid contamination.

Key Tips

• Always check the lot-specific datasheet for any unique instructions.
• Avoid vigorous mixing or vortexing.
• Use low endotoxin reagents and buffers to minimize background effects in cell culture.
• Aliquot and freeze unused portions to minimize freeze-thaw cycles.

By following these steps, you will ensure proper reconstitution, stability, and bioactivity of Recombinant Mouse IL-20 for your cell culture experiments.