Recombinant Mouse IL-13 Rα2

Recombinant Mouse IL-13 Rα2 is a valuable tool in research applications focused on immunology, inflammation, and cancer biology due to its role as a high-affinity decoy receptor for IL-13 and its involvement in modulating type 2 immune responses.

Key scientific applications and rationale:

• Modulation of IL-13 Signaling: IL-13Rα2 binds IL-13 with much higher affinity than IL-13Rα1/IL-4Rα, acting as a physiological rheostat that limits IL-13-driven STAT6 signaling and type 2 immunity. Using recombinant IL-13Rα2 allows you to experimentally regulate IL-13 bioavailability and downstream effects in cell-based or animal models.

• Inflammatory Disease Models: In murine models of inflammatory bowel disease (IBD), IL-13Rα2 slows recovery by restricting the anti-inflammatory functions of IL-13. Neutralizing or supplementing IL-13Rα2 can help dissect its role in disease progression and tissue repair, making it useful for mechanistic studies and therapeutic screening.

• Cancer Research: IL-13Rα2 is overexpressed in various tumors and can promote tumor proliferation, survival, invasion, and metastasis. Recombinant IL-13Rα2 is used to study tumor biology, test targeted therapies (e.g., CAR T cells, antibody-drug conjugates), and evaluate the specificity and efficacy of IL-13Rα2-directed interventions.

• Cellular Assays: Recombinant mouse IL-13Rα2 can inhibit IL-13-induced proliferation in certain cell lines (e.g., TF-1 erythroleukemic cells), providing a functional readout for IL-13 activity and receptor interactions.

• Therapeutic Target Validation: By adding recombinant IL-13Rα2 to in vitro or in vivo systems, researchers can validate its role as a therapeutic target, assess its impact on cytokine networks, and explore its potential in modulating immune responses or tumor microenvironments.

Best practices:

• Use recombinant IL-13Rα2 in dose-response studies to determine its effect on IL-13 signaling and cellular outcomes.
• Combine with other cytokines or receptor antagonists to dissect pathway specificity.
• Employ in disease models (e.g., colitis, cancer xenografts) to evaluate therapeutic potential and mechanistic effects.

Summary:
Recombinant Mouse IL-13 Rα2 is essential for investigating the regulation of IL-13-mediated immune responses, understanding its dual role in inflammation and cancer, and validating therapeutic strategies targeting the IL-13/IL-13Rα2 axis.

Recombinant Mouse IL-13 Rα2 can be used as a standard for quantification or calibration in ELISA assays specifically designed to measure IL-13 Rα2, but not for assays targeting IL-13 itself. The suitability depends on the assay's intended analyte and validation.

• For quantifying IL-13 Rα2:
  Recombinant Mouse IL-13 Rα2 is routinely used as a standard in ELISA kits designed to measure IL-13 Rα2 concentrations in biological samples such as cell culture supernatants, tissue homogenates, serum, and plasma. These kits include recombinant IL-13 Rα2 as the calibrator to generate a standard curve, enabling accurate quantification of IL-13 Rα2 in unknown samples.

• For quantifying IL-13:
  Recombinant IL-13 Rα2 should not be used as a standard for IL-13 quantification. ELISA kits for IL-13 use recombinant IL-13 as the standard, and the presence of IL-13 Rα2 (as a binding protein) can potentially interfere with IL-13 detection if not properly controlled. Studies have shown that recombinant IL-13 Rα2 does not significantly interfere with IL-13 quantification under certain assay conditions, but it is not suitable as a standard for IL-13 measurement.

• Assay validation:
  When using recombinant proteins as standards, it is critical to ensure that the protein's purity, concentration, and biological activity are well-characterized. The standard should match the analyte detected by the assay in terms of epitope recognition and molecular form. ELISA kits validated for IL-13 Rα2 quantification provide recombinant IL-13 Rα2 as the standard, and protocols specify its use for calibration.

• Best practices:
  Always use the standard recommended by the ELISA kit manufacturer for the specific analyte being measured. If developing a custom ELISA, validate the recombinant IL-13 Rα2 standard for accuracy, precision, and parallelism with endogenous samples.

Summary Table:

Do not use recombinant IL-13 Rα2 as a standard for IL-13 quantification. Use it only for assays specifically measuring IL-13 Rα2.

Recombinant Mouse IL-13 Rα2 has been validated in published research for several key applications, primarily in cancer biology, immunotherapy development, and mechanistic studies of cytokine signaling.

Validated Applications:

• Cancer Immunotherapy Research:
  Recombinant Mouse IL-13 Rα2 is widely used to study and validate chimeric antigen receptor (CAR) T cell therapies targeting IL-13Rα2-expressing tumors, including glioblastoma, melanoma, ovarian carcinoma, and colorectal cancer. It serves as a target antigen in cytotoxicity assays, cytokine secretion assays, and in vivo mouse xenograft models to assess CAR T cell specificity, efficacy, and trafficking.

• Signaling Pathway Analysis:
  It is employed to dissect IL-13Rα2-mediated signaling pathways, including its role as a decoy receptor and its involvement in tumor proliferation, invasion, and metastasis. Studies use recombinant IL-13 Rα2 to investigate downstream effects such as apoptosis, cell survival, and tissue responses, often in combination with other proteins or genetic models.

• Therapeutic Peptide and Antibody Validation:
  Recombinant IL-13 Rα2 is used to validate the binding and inhibitory activity of therapeutic peptides and antibodies designed to block IL-13Rα2 function, both in vitro (migration, invasion, proliferation assays) and in vivo (mouse survival and metastasis models).

• Receptor-Ligand Interaction Studies:
  It is utilized in binding assays to characterize the affinity and selectivity of engineered IL-13 variants, muteins, and immunotoxins for IL-13Rα2 versus IL-13Rα1, informing the design of more selective therapeutic agents.

• Mechanistic Studies in Allergy and Inflammation:
  Recombinant IL-13 Rα2 is used to explore its role in airway hyperresponsiveness and allergic asthma, as well as its function in modulating immune cell infiltration and cytokine production in lung injury models.

Common Experimental Protocols:

• Flow cytometry for receptor expression validation.
• Cytotoxicity and cytokine secretion assays in co-culture systems.
• In vivo mouse models for tumor growth, metastasis, and survival analysis.
• ELISA and Luminex assays for cytokine quantification.
• Peptide and antibody binding assays for therapeutic validation.

Summary Table:

These applications demonstrate the versatility of recombinant Mouse IL-13 Rα2 in both basic and translational research, particularly in the context of cancer immunotherapy and cytokine signaling.

To reconstitute and prepare Recombinant Mouse IL-13 Rα2 protein for cell culture experiments, first centrifuge the vial briefly to collect all lyophilized material at the bottom, then add sterile buffer (commonly PBS or Tris-based buffer) to achieve your desired concentration, typically 50–100 µg/mL, and include a carrier protein such as 0.1% BSA or HSA for stability.

Detailed protocol:

• Centrifuge the vial for 20–30 seconds to ensure all powder is at the bottom.
• Reconstitute using sterile buffer. For most recombinant proteins, sterile PBS or Tris buffer (pH 7.2–7.5) is suitable. Some preparations may be supplied in Tris/glycine/trehalose and can be reconstituted in sterile water or buffer as specified by the datasheet.
• Add carrier protein: For long-term stability and to prevent adsorption to plastic, add 0.1% BSA or HSA to the buffer.
• Typical concentration: Prepare a stock solution at 50–100 µg/mL. Adjust as needed for your experimental design.
• Mix gently by pipetting up and down or gentle vortexing. Avoid vigorous agitation to prevent protein denaturation.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
• Storage: Store aliquots at –20°C or colder. For short-term use (days to weeks), 4°C is acceptable if carrier protein is present.

Example reconstitution (for 100 µg vial):

• Add 1 mL sterile PBS with 0.1% BSA to achieve 100 µg/mL stock.
• Mix gently until fully dissolved.

Additional notes:

• Always consult the specific product datasheet for any unique requirements.
• Confirm protein integrity by SDS-PAGE if needed.
• For cell culture, ensure all solutions are sterile and endotoxin levels are appropriate for your application.

This protocol is broadly applicable to recombinant IL-13 Rα2 proteins used in cell culture, but always verify with the product-specific documentation for optimal results.