Recombinant Human IL-22

Recombinant Human IL-22 is widely used in research because it is a key cytokine involved in epithelial tissue protection, regeneration, and immune modulation, making it valuable for studying tissue repair, inflammation, and disease models involving barrier organs such as the gut, lung, liver, and skin.

Key scientific reasons to use Recombinant Human IL-22 in research applications:

• Tissue Protection and Regeneration: IL-22 promotes epithelial cell survival, proliferation, and regeneration after injury or infection, especially at barrier surfaces (e.g., gut, lung, skin, liver, kidney). For example, recombinant IL-22 enhances recovery of intestinal stem cells and epithelial regeneration after bone marrow transplantation, reducing pathology and mortality in graft-versus-host disease models.
• Disease Modeling and Therapeutic Potential: IL-22 is implicated in the pathogenesis and potential treatment of various non-autoimmune diseases, including acute lung injury, atherosclerosis, fatty liver disease, and kidney injury. Its ability to reduce liver triglycerides and control lipid metabolism via STAT3 signaling is particularly relevant for metabolic and liver disease research.
• Unique Signaling Pathways: IL-22 signals through the IL-22R/IL-10R2 complex, activating JAK-STAT and ERK pathways, which are distinct from other cytokines like IL-10, allowing for the dissection of specific signaling mechanisms in epithelial and immune cells.
• Immunomodulation: IL-22 modulates immune responses at mucosal surfaces, influencing inflammation, barrier integrity, and host defense against pathogens. It is upregulated in many chronic inflammatory diseases, making it a target for studying immune regulation and pathology.
• Bioassays and Functional Studies: Recombinant Human IL-22 is used in cell-based assays to study cytokine signaling, epithelial cell biology, and immune cell interactions, as well as in organoid cultures and animal models to assess its effects on tissue repair and disease outcomes.

Additional applications include:

• Investigating the role of IL-22 in autoimmune and infectious diseases.
• Exploring its effects on stem cell biology and organoid growth.
• Testing therapeutic strategies for tissue injury and inflammatory disorders.

In summary, Recombinant Human IL-22 is a versatile tool for research into epithelial biology, tissue repair, immune modulation, and disease mechanisms, with broad applications in both basic and translational studies.

Yes, recombinant human IL-22 can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and properly validated for this purpose. This is a common practice in cytokine ELISA protocols, where recombinant proteins serve as reference standards to generate calibration curves for quantifying endogenous IL-22 in biological samples.

Supporting details:

• ELISA kits for human IL-22 routinely use recombinant human IL-22 as the standard. Technical datasheets and protocols specify that both natural and recombinant IL-22 are accurately quantified using these assays, and standard curves are generated using recombinant IL-22.
• Validation and purity are critical. The recombinant IL-22 standard should be well-characterized, with confirmed identity, purity, and biological activity. It should be free of contaminants that could interfere with antibody binding or detection.
• Standard curve preparation: The recombinant IL-22 is serially diluted in the recommended assay buffer or diluent to create a standard curve, typically spanning the assay’s dynamic range (e.g., 0.78–50 pg/mL or higher, depending on kit sensitivity).
• Parallelism: ELISA kits often demonstrate parallelism between curves generated from recombinant standards and those from endogenous IL-22 in biological samples, confirming the suitability of recombinant IL-22 for calibration.
• Matrix compatibility: Recombinant IL-22 standards are compatible with various sample matrices, including serum, plasma, and tissue culture media, as long as the assay is validated for those matrices.

Best practices:

• Use the recombinant IL-22 standard supplied or recommended by the ELISA kit manufacturer, as it will have been validated for use with the specific antibodies and assay conditions.
• If using a recombinant IL-22 from another source, ensure it matches the sequence and post-translational modifications (if relevant) of endogenous human IL-22, and verify its performance in your assay by running parallelism and recovery experiments.
• Store and handle the recombinant standard according to manufacturer instructions to maintain stability and activity.

Limitations:

• If your recombinant IL-22 differs significantly in structure or glycosylation from endogenous IL-22, or if it contains tags or fusion partners, it may affect antibody recognition and quantification accuracy. Always confirm equivalence through assay validation.
• High concentrations of IL-22 binding protein (IL-22BP) in samples may interfere with detection of free IL-22, affecting quantification.

Summary: Recombinant human IL-22 is widely accepted and validated as a standard for ELISA quantification, provided it is properly characterized and matched to the assay system.

Recombinant Human IL-22 has been validated for a broad range of applications in published research, primarily in studies involving cell-based bioassays, cell culture, and functional analyses of epithelial and immune responses.

Key validated applications include:

• Bioassays: Used to assess IL-22 activity, such as induction of downstream cytokines (e.g., IL-10 secretion in COLO 205 cells), activation of signaling pathways (e.g., STAT3), and modulation of cellular responses in various cell types including keratinocytes, hepatocytes, and immune cells.
• Cell Culture: Applied to study effects on epithelial cells, immune cell differentiation, and tissue-specific responses, including barrier integrity, proliferation, and wound healing.
• Surface Plasmon Resonance: Used to characterize protein-protein interactions and binding kinetics involving IL-22 and its receptor.
• Ligand Blocking: Utilized to block IL-22 receptor interactions in specificity assays, confirming functional activity and receptor engagement.
• In Vivo Models: Recombinant IL-22 and IL-22 fusion proteins have been tested in animal models for therapeutic potential in conditions such as colitis, hepatitis, pneumonitis, acute respiratory distress syndrome (ARDS), and epithelial injury.
• Clinical Research: Recombinant IL-22-Fc fusion proteins have entered clinical trials for treatment of gastrointestinal graft-versus-host disease (GI-GVHD) and ARDS, demonstrating safety and efficacy in modulating epithelial inflammation and repair.

Representative published research applications:

• Immunomodulation: Studying suppression or activation of immune cell subsets (e.g., Th17, regulatory T cells) in autoimmune diseases such as rheumatoid arthritis and multiple sclerosis.
• Epithelial Barrier Function: Investigating enhancement of tight junctions, mucus production, and antimicrobial peptide expression in epithelial tissues, relevant to host defense and tissue regeneration.
• Cancer Research: Assessing synergistic effects with other cytokines (e.g., IL-6, TNF-α) on cancer cell growth and signaling pathways.
• Wound Healing and Tissue Repair: Evaluating keratinocyte proliferation, VEGF production, and tissue remodeling in cutaneous wound models.
• Host Defense: Exploring innate immune responses against bacterial pathogens in epithelial cells, including lung and gastrointestinal tract.

Summary Table of Validated Applications

These applications demonstrate that recombinant human IL-22 is a versatile tool for investigating epithelial biology, immune modulation, and therapeutic interventions in both preclinical and clinical research settings.

To reconstitute and prepare Recombinant Human IL-22 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS), ideally supplemented with 0.1% carrier protein such as human or bovine serum albumin (BSA), to a working concentration appropriate for your assay.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile PBS (pH 7.2–7.4) to the vial. A common reconstitution concentration is 10–100 μg/mL depending on the manufacturer’s recommendation and your experimental needs.
  • For carrier-free formulations, add 0.1% BSA to the PBS to minimize protein adsorption and loss.
• Gently mix by inverting or swirling. Avoid vigorous vortexing or pipetting to prevent protein denaturation or foaming.
• Allow the solution to sit at room temperature for 15–30 minutes to ensure complete dissolution.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at −20 °C or below for long-term storage. For short-term use (up to 2–7 days), store at 4 °C.

Example calculation for a 10 μg/mL stock:If your vial contains 50 μg IL-22, add 5 mL sterile PBS + 0.1% BSA to achieve 10 μg/mL.

Additional notes:

• If your application is highly sensitive to endotoxin, confirm the endotoxin level is suitable for cell culture (typically <0.1 EU/μg).
• For functional assays, confirm the biological activity using a relevant cell-based assay (e.g., induction of IL-10 secretion in COLO-205 cells).
• Always consult the product-specific datasheet for any unique instructions, as formulations may vary.

Summary of best practices:

• Use sterile technique throughout.
• Use PBS (pH 7.2–7.4), with 0.1% BSA if carrier-free.
• Avoid repeated freeze-thaw cycles.
• Aliquot and store appropriately.

These steps will ensure optimal solubility, stability, and biological activity of recombinant human IL-22 for cell culture experiments.