Recombinant Mouse IL-10

Using recombinant Mouse IL-10 in research applications is essential for studying immune regulation, inflammation, and disease models because IL-10 is a key anti-inflammatory cytokine with well-characterized immunomodulatory functions.

Key reasons to use recombinant Mouse IL-10:

• Immunosuppression and Regulation: IL-10 is a potent suppressor of effector T cell proliferation and cytokine production, and it is crucial for the function of regulatory T cells. It downregulates Th1 cytokines, MHC class II antigens, and costimulatory molecules, thereby limiting excessive immune responses and preventing tissue damage.
• B Cell Support: IL-10 enhances B cell survival, proliferation, and antibody production, making it valuable for studies on humoral immunity and B cell biology.
• Anti-inflammatory Effects: IL-10 blocks pro-inflammatory signaling pathways such as NF-κB and modulates the JAK-STAT pathway, helping to resolve inflammation and maintain immune homeostasis.
• Disease Modeling: Recombinant Mouse IL-10 is widely used in models of autoimmune disease, infection, cancer, and tissue injury to dissect its role in disease progression and therapy. For example, it has been shown to enhance the effectiveness of cancer vaccines and suppress inflammation in various in vivo models.
• Reproducibility and Standardization: Recombinant proteins provide consistent, defined activity and purity, which is critical for reproducible results in cell culture, functional assays, and in vivo studies.
• Therapeutic Potential: Engineered forms of recombinant IL-10 with improved stability and efficacy are being developed as potential anti-inflammatory and immunotherapeutic agents.

Common applications include:

• Cell culture experiments to study immune cell differentiation and function.
• In vivo models to investigate the effects of IL-10 on inflammation, autoimmunity, infection, and cancer.
• Functional assays to assess cytokine signaling, immune modulation, and therapeutic interventions.

In summary, recombinant Mouse IL-10 is a versatile tool for dissecting immune mechanisms, modeling disease, and developing new therapeutic strategies due to its central role in immune regulation and anti-inflammatory responses.

Yes, recombinant mouse IL-10 can be used as a standard for quantification or calibration in ELISA assays, provided it is suitable for your specific assay format and detection system.

Key Points:

• Purpose: Recombinant mouse IL-10 is commonly used as a quantitative standard in ELISA assays to generate a standard curve, which allows for the accurate measurement of IL-10 concentrations in experimental samples.
• Purity and Endotoxin: High-purity recombinant mouse IL-10 (≥95% pure by SDS-PAGE, low endotoxin levels) is recommended for reliable results. For example, products from reputable suppliers such as BD Pharmingen, R&D Systems, BioLegend, and eBioscience are specifically formulated for use as ELISA standards.
• Standard Curve: To ensure accurate quantification, prepare a dilution series of the recombinant IL-10 standard (typically doubling dilutions, e.g., from 2,000 pg/mL down to 15 pg/mL) and include it on each ELISA plate.
• Sample Compatibility: The standard curve generated with recombinant IL-10 can be used to quantify both recombinant and natural mouse IL-10 in samples such as cell culture supernatants, serum, and plasma, as long as the assay antibodies recognize both forms.
• Carrier Protein: For optimal stability and activity, it is recommended to use carrier proteins (e.g., BSA) at appropriate concentrations (5–10 mg/mL for ELISA standards, 1–2 mg/mL for biological assays). Avoid diluting the standard below 50 µg/mL for long-term storage.
• Kit Compatibility: Many commercial ELISA kits (e.g., BD OptEIA, R&D Systems Quantikine, BioLegend ELISA MAX, eBioscience Ready-Set-Go!) are designed to work with recombinant mouse IL-10 standards and provide detailed protocols for reconstitution and dilution.

Recommendations:

• Always follow the manufacturer's instructions for reconstitution and dilution of the recombinant IL-10 standard.
• Validate the standard curve in your specific assay conditions to ensure linearity and accuracy.
• Use appropriate controls and calibrators to confirm the reliability of your quantification.

In summary, recombinant mouse IL-10 is a suitable and widely used standard for quantification and calibration in ELISA assays, provided it meets the purity and formulation requirements for your experimental setup.

Recombinant Mouse IL-10 has been validated for a range of applications in published research, primarily in bioassays, in vivo studies, and as a standard in immunoassays (ELISA).

Key validated applications include:

• Bioassays:

  • Used to assess biological activity in cell-based systems, such as proliferation assays with the MC/9 cell line.
  • Studied for its effects on immune cell differentiation, macrophage polarization, and cytokine production in vitro.
  • Investigated in mechanistic studies of signaling pathways, such as inhibition of oxidative stress-induced activation of p38 and NF-κB in vascular cells.

• In Vivo Studies:

  • Administered to mice to evaluate its therapeutic effects in models of inflammation, autoimmune disease, atherosclerosis, pulmonary fibrosis, and cancer.
  • Used to modulate immune responses in models of food allergy, viral infection, and neurodegenerative disease.
  • Applied as an adjuvant to enhance the effectiveness of cancer vaccines and to study its impact on tumor growth and metastasis.

• Immunoassays (ELISA):

  • Employed as a standard for quantifying IL-10 levels in biological samples.

• Functional and Differentiation Assays:

  • Used in studies of T cell and macrophage function, including regulatory T cell induction and macrophage polarization.
  • Applied in protocols for cell culture and differentiation studies to investigate anti-inflammatory and immunoregulatory mechanisms.

Additional validated uses in published research:

• Suppression of pro-inflammatory responses and enhancement of B cell proliferation and immunoglobulin secretion.
• Inhibition of antigen presentation by monocytes and macrophages, including downregulation of MHC class II and costimulatory molecules.
• Investigation of IL-10’s role in infection, autoimmunity, and neuroinflammation.

These applications are supported by numerous peer-reviewed studies, with recombinant mouse IL-10 cited in over 70 publications for these purposes. The protein is widely used as a tool to dissect immunoregulatory pathways and to model therapeutic interventions in preclinical research.

Reconstitution Guidelines

Recombinant Mouse IL-10 is typically supplied in lyophilized form and requires proper reconstitution before use in cell culture experiments. The standard reconstitution concentration is 100 μg/mL in sterile PBS, though some protocols recommend reconstituting at 0.1 mg/mL (100 μg/mL) with sterile water.

Reconstitution Buffer Selection

The choice of reconstitution buffer depends on the formulation variant:

• With BSA carrier protein: Reconstitute in sterile PBS containing at least 0.1% human or bovine serum albumin
• With Trehalose: Reconstitute in sterile PBS without additional carrier protein
• Alternative approach: Some formulations can be reconstituted with sterile water at 0.1 mg/mL and subsequently diluted into aqueous solutions as needed

Preparation Protocol

Step 1: Initial ReconstitutionAdd the appropriate volume of reconstitution buffer to the lyophilized protein vial according to the manufacturer's specifications. Allow the reconstituted standard to sit for approximately 15 minutes at room temperature, then mix gently prior to making dilutions.

Step 2: Sample Preparation for Cell CultureFor cell culture supernatant samples, centrifuge at 2,000 × g for 10 minutes to remove debris before analysis. If using cell lysates, centrifuge at 1,000 × g for 5 minutes and wash cells three times with pre-cooled PBS.

Storage and Stability

Proper storage is critical for maintaining protein activity:

• Short-term storage: Up to 1 month at 2-8°C under sterile conditions after reconstitution
• Long-term storage: 3 months at -20 to -70°C under sterile conditions after reconstitution
• Critical consideration: Use a manual defrost freezer and avoid repeated freeze-thaw cycles

When storing reconstituted protein, prepare working aliquots to minimize freeze-thaw exposure. For cell culture supernatant and serum samples, store at less than -20°C and similarly avoid repeated freeze-thaw cycles.

Bioactivity Considerations

The recombinant protein is typically supplied as a 17-18.8 kDa monomer that functions as a noncovalently associated homodimer in biological systems. For cell proliferation assays using mast cell lines, the effective dose (ED₅₀) ranges from 0.1-0.6 ng/mL, providing a reference for appropriate working concentrations.