Recombinant Mouse IL-9

Recombinant Mouse IL-9 is used in research to study its diverse roles in immune regulation, inflammation, allergy, tumor immunity, and cell signaling, making it a valuable tool for dissecting cytokine-driven mechanisms in mouse models.

Key applications and scientific rationale include:

• Allergic Inflammation and Asthma Models: IL-9 promotes the expansion and recruitment of mast cells and eosinophils, enhances IgE and Th2 cytokine production, and is genetically linked to asthma susceptibility. Recombinant IL-9 can be used to induce or modulate allergic responses in vivo and in vitro, allowing researchers to study mechanisms of airway hyperresponsiveness and anaphylaxis.

• Mast Cell Biology: IL-9 is a potent growth factor for mast cell progenitors, driving their proliferation and tissue accumulation during allergic inflammation. Recombinant IL-9 enables controlled studies of mast cell development, migration, and function in mouse systems.

• Parasitic Infection and Mucosal Immunity: IL-9 enhances mucosal mast cell responses that facilitate parasite expulsion from the gastrointestinal tract, making it useful for modeling host defense mechanisms against helminths.

• Tumor Immunology and Adoptive Cell Therapy: IL-9 modulates CD8+ T cell cytotoxicity and can enhance anti-tumor immune responses. Recombinant IL-9 is used to investigate its effects on T cell function, tumor infiltration, and the development of memory-like T cell phenotypes in cancer models.

• Autoimmunity and Immune Regulation: IL-9 has context-dependent pro- and anti-inflammatory effects, influencing Treg-mediated suppression and macrophage polarization. It is used to study immune modulation in autoimmune disease models.

• Cell Signaling Studies: IL-9 signals through the IL-9 receptor (IL9R), activating STAT pathways and affecting cell proliferation and survival. Recombinant IL-9 is essential for dissecting these signaling cascades in primary cells or cell lines.

• Therapeutic Target Validation: Because IL-9 is implicated in both pathogenic and protective immune responses, recombinant protein is used to validate therapeutic strategies targeting the IL-9/IL-9R axis in preclinical models.

In summary, recombinant mouse IL-9 is a critical reagent for immunological research, enabling precise manipulation of IL-9-dependent pathways in mouse systems to elucidate its roles in health and disease.

Yes, recombinant mouse IL-9 can be used as a standard for quantification and calibration in ELISA assays, but with important considerations regarding the specific formulation and intended application.

Appropriate Use Cases

Recombinant mouse IL-9 proteins with bovine serum albumin (BSA) formulations are specifically recommended for use as ELISA standards. These standards are suitable for establishing standard curves and performing quantitative measurements of natural and recombinant mouse IL-9 in sandwich ELISA formats.

Critical Limitations

However, there is a significant caveat: ELISA standard recombinant proteins are not recommended for bioassay applications, as they are not validated for these uses. Additionally, if you are using bioassay-grade standards, the mass values assigned to these proteins describe potency in bioassay only and should not be used for immunoassay quantification, as the mass assignments are not absolute values.

Protein Characteristics to Consider

When selecting a recombinant mouse IL-9 standard, be aware of the protein composition. Some recombinant formulations are non-glycosylated polypeptides produced in bacterial expression systems, whereas native mouse IL-9 is heavily glycosylated. This structural difference may affect how the recombinant protein behaves in your assay compared to naturally occurring IL-9.

Recommendation

For ELISA quantification purposes, use recombinant mouse IL-9 standards that are specifically formulated and validated for immunoassay applications. Verify that your chosen standard is appropriate for ELISA use rather than bioassay applications, and ensure the formulation includes appropriate carriers like BSA if you plan to use it in cell or tissue culture applications or as an ELISA standard.

Recombinant Mouse IL-9 has been validated in published research for a range of applications, primarily in immunological and cell biology studies involving both in vitro and in vivo models.

Key validated applications include:

• In vivo functional studies: Recombinant Mouse IL-9 (rIL-9) is widely used to investigate its biological effects in mouse models. These include:

  • Mucosal immunity and helminth expulsion: rIL-9 administration enhances mucosal mast cell responses, promoting worm expulsion in models of parasitic infection.
  • Allergic inflammation and asthma models: rIL-9 is used to exacerbate or modulate mast cell-dependent allergic inflammation, airway hyperresponsiveness, and systemic anaphylaxis.
  • Tumor immunology: rIL-9 has been used to modulate anti-tumor immunity, particularly by enhancing CD8+ T cell cytotoxicity and supporting adoptive cell therapy approaches.
  • Autoimmune and neuroinflammatory disease models: rIL-9 has been shown to reduce inflammation and protect from neurodegeneration in mouse models of multiple sclerosis.

• In vitro cell culture assays:

  • Cell proliferation and differentiation: rIL-9 is validated for inducing proliferation in cell lines such as MO7e human megakaryocytic leukemic cells.
  • B cell and T cell functional assays: rIL-9 is used to stimulate B cells (enhancing IgG and IgE production) and T cells (modulating cytokine production and proliferation).
  • Macrophage and monocyte polarization: rIL-9 is used to induce anti-inflammatory phenotypes in monocytes and macrophages, including increased TGFβ production and reduced pro-inflammatory cytokine release.
  • Eosinophil maturation and survival: rIL-9 enhances eosinophil precursor maturation and inhibits eosinophil apoptosis in vitro.

• Adoptive cell therapy and CAR-T cell engineering: Recombinant Mouse IL-9 has been used in studies engineering T cells with synthetic or chimeric IL-9 receptors to enhance anti-tumor efficacy in mouse models.

• Functional validation in immunological assays: rIL-9 is used to study its effects on regulatory T cells (Tregs), mast cells, and innate lymphoid cells (ILC2), including their proliferation, survival, and cytokine production.

Summary Table: Validated Applications of Recombinant Mouse IL-9

These applications are supported by multiple peer-reviewed studies and reviews, confirming the broad utility of recombinant Mouse IL-9 in immunological research.

To reconstitute and prepare Recombinant Mouse IL-9 protein for cell culture experiments, dissolve the lyophilized protein in sterile distilled water or sterile buffer to a concentration of 0.1–1.0 mg/mL. For optimal stability and activity, further dilute in an aqueous buffer containing a carrier protein such as 0.1–1.0% BSA (bovine serum albumin) or HSA (human serum albumin).

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile distilled water or buffer (e.g., PBS) to achieve a concentration between 0.1–1.0 mg/mL, depending on your experimental requirements.
• Gently pipette to dissolve the protein completely, washing down the sides of the vial to recover all material.
• For working solutions, dilute the reconstituted stock in cell culture medium or buffer containing 0.1–1.0% BSA or HSA to prevent adsorption and loss of activity.
• Aliquot the stock solution to avoid repeated freeze-thaw cycles, which can denature the protein.
• Storage: Store lyophilized protein at –20°C or lower. After reconstitution, aliquots can be stored at –20°C to –80°C for long-term use, or at 2–8°C for up to one week.

Additional notes:

• The optimal working concentration for cell culture should be determined empirically for your specific assay.
• Avoid repeated freeze-thaw cycles; always use fresh aliquots for each experiment.
• If the protein is provided carrier-free, always add carrier protein for storage and dilution to maintain stability.

This protocol ensures maximal recovery, stability, and biological activity of recombinant mouse IL-9 for cell culture applications.