Recombinant Mouse IL-10

Recombinant Mouse IL-10 is widely used in research because it is a potent anti-inflammatory cytokine with critical roles in immune regulation, making it valuable for studies on immune modulation, inflammation, autoimmunity, and tissue repair.

Key reasons to use recombinant Mouse IL-10 in research applications:

• Immunosuppression and Anti-Inflammatory Effects: IL-10 inhibits the activation and effector functions of various immune cells, including macrophages and T cells, thereby suppressing pro-inflammatory cytokine production and limiting tissue damage during immune responses. This makes it essential for studying mechanisms of immune tolerance, inflammation, and resolution of immune-mediated diseases.

• Regulation of T Regulatory Cells: IL-10 is crucial for the function of regulatory T cells (Tregs), which are central to maintaining immune homeostasis and preventing autoimmunity by suppressing effector T cell proliferation and cytokine production.

• B Cell Modulation: IL-10 enhances B cell survival, proliferation, and antibody production, making it relevant for research on humoral immunity and autoimmune diseases.

• Modeling Disease and Therapeutic Interventions: Recombinant Mouse IL-10 is used in models of autoimmune diseases, inflammatory disorders, and cancer to investigate its therapeutic potential and underlying mechanisms. For example, it has been shown to attenuate tissue remodeling in cardiovascular disease models and enhance anti-tumor immune responses in cancer models.

• Cell Culture and Bioassays: Recombinant IL-10 is used as a standard in ELISA, as a control in Western blots, and to stimulate or inhibit specific cell types in vitro, ensuring reproducibility and precision in immunology and cell biology experiments.

• Mechanistic Studies: IL-10 is involved in blocking NF-κB activity and regulating the JAK-STAT signaling pathway, which are central to many immune and inflammatory processes.

• Improved Stability and Activity: Engineered recombinant forms of IL-10, such as stable dimers, have been developed with enhanced biological activity and stability, further expanding its utility in research and potential therapeutic applications.

In summary, recombinant Mouse IL-10 is a versatile tool for dissecting immune regulatory pathways, modeling disease, and testing therapeutic strategies targeting inflammation and immune responses.

Yes, recombinant mouse IL-10 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly formulated and validated for this purpose. Multiple commercial ELISA kits and protocols specifically use recombinant mouse IL-10 as the standard for generating calibration curves.

Key considerations and supporting details:

• Intended Use: Recombinant mouse IL-10 is widely used as a quantitative standard in ELISA assays to measure IL-10 levels in biological samples such as cell culture supernatants, serum, and plasma.
• Formulation: For use as an ELISA standard, it is recommended to prepare the recombinant protein in a carrier protein solution (e.g., BSA) at concentrations of 5–10 mg/mL to maintain stability and prevent adsorption to plasticware. Avoid diluting below 50 µg/mL for long-term storage.
• Validation: Commercial ELISA kits demonstrate that recombinant mouse IL-10 produces standard curves that are parallel to those generated with natural mouse IL-10, indicating equivalence for quantification purposes. This ensures accurate and reliable quantification of both recombinant and endogenous IL-10.
• Preparation: Follow the specific reconstitution and dilution instructions provided by your ELISA kit or protocol. Typically, a series of doubling dilutions is used to generate a standard curve covering the expected range of IL-10 concentrations in your samples.
• Carrier Protein: The choice of carrier protein (e.g., BSA) should be pre-screened for compatibility with your assay system, as some carriers may interfere with antibody binding or detection.
• Storage: Store reconstituted standards in aliquots at -70 °C or as recommended to avoid repeated freeze-thaw cycles, which can degrade protein activity.

Limitations:

• Ensure the recombinant IL-10 standard is compatible with the antibodies and detection system used in your specific ELISA assay. Some kits are optimized for their own matched antibody pairs and diluents.
• For absolute quantification, the recombinant standard should be of high purity and its concentration accurately determined.

Summary Table: Use of Recombinant Mouse IL-10 as ELISA Standard

In summary: Recombinant mouse IL-10 is a validated and widely accepted standard for ELISA quantification, provided you follow best practices for preparation, storage, and compatibility with your assay system.

Recombinant Mouse IL-10 has been validated for a broad range of applications in published research, including bioassays, in vivo studies, ELISA standards, cell culture, differentiation studies, and functional immunological assays.

Key validated applications include:

• Bioassays: Used to assess IL-10’s biological activity, such as its ability to suppress pro-inflammatory cytokine production, regulate immune cell differentiation (e.g., regulatory T cells, macrophage polarization), and modulate signaling pathways like NF-κB and JAK-STAT.
• In Vivo Studies: Applied in mouse models to study IL-10’s effects on disease processes, including attenuation of aortic remodeling, reversal of food allergen sensitivity, suppression of autoimmune encephalomyelitis, modulation of pulmonary fibrosis, and enhancement of anti-tumor immunity.
• ELISA Standard: Used as a standard protein in ELISA assays to quantify IL-10 levels in biological samples.
• Cell Culture and Differentiation: Employed to study B cell survival, proliferation, antibody production, and differentiation of immune cell subsets in vitro.
• Functional Immunological Assays: Investigated for its role in suppressing antigen presentation, downregulating MHC class II and costimulatory molecules, and controlling inflammatory responses.

Additional validated uses include:

• Adjuvant in Vaccine Studies: Shown to enhance the effectiveness of recombinant anti-cancer vaccines and augment T-cell-mediated responses in murine models.
• Pathogenesis and Disease Models: Used to explore IL-10’s regulatory role in infection, autoimmunity, and neurodegenerative disease models (e.g., synucleinopathy, lupus, rheumatoid arthritis).

Summary Table: Validated Applications of Recombinant Mouse IL-10

These applications are supported by numerous peer-reviewed publications, demonstrating the protein’s utility in both basic and translational immunology research.

To reconstitute and prepare Recombinant Mouse IL-10 protein for cell culture experiments, dissolve the lyophilized protein in sterile water or buffer to a recommended stock concentration, then dilute as needed for your assay. The following protocol summarizes best practices based on multiple authoritative sources:

1. Reconstitution

• Add sterile water to the lyophilized IL-10 to achieve a concentration of 0.1 mg/mL (100 μg/mL).
• Alternatively, some protocols recommend using sterile PBS (phosphate-buffered saline), especially if the product contains stabilizers like BSA or trehalose.
• If using PBS, it is advisable to include at least 0.1% carrier protein (such as BSA or HSA) to minimize adsorption and loss of activity.

2. Mixing

• Gently swirl or invert the vial to dissolve the protein completely.
• Avoid vigorous vortexing or pipetting, which may denature the protein.

3. Aliquoting and Storage

• After reconstitution, aliquot the solution into small volumes to avoid repeated freeze-thaw cycles.
• Store aliquots at –20°C or –70°C for long-term storage; for short-term use (up to 1 week), store at 2–8°C.
• Avoid repeated freeze-thaw cycles, as this can reduce protein activity.

4. Working Solution Preparation

• Before use in cell culture, dilute the stock solution to the desired working concentration using cell culture medium or sterile PBS with carrier protein if needed.
• Typical working concentrations for cell stimulation assays range from 0.1–10 ng/mL, but optimal concentrations should be determined empirically for your specific application.

5. Additional Notes

• Ensure all solutions and diluents are sterile and endotoxin-free, especially for sensitive cell culture applications.
• If the protein is to be used in functional assays, confirm activity with a bioassay or reference standard if possible.

Summary Table

These guidelines are consistent with standard protocols for recombinant cytokines and specifically for mouse IL-10. Always consult the specific product datasheet for any manufacturer-specific recommendations.