Recombinant Mouse IL-1 RI

Recombinant mouse IL-1RI offers several compelling advantages for research applications, particularly in studies examining interleukin-1 signaling pathways and inflammatory responses.

Key Research Applications

Pathway Characterization and Mechanistic Studies

Recombinant mouse IL-1RI enables detailed investigation of IL-1 receptor signaling mechanisms. This protein is particularly valuable for studying how IL-1 signaling regulates immune cell differentiation and function. For instance, IL-1RI plays a critical role in Th17 cell differentiation through IRF4-dependent mechanisms, making it essential for research into autoimmune diseases like multiple sclerosis. By using the recombinant receptor, you can isolate and characterize specific signaling events without the complexity of whole-cell systems.

Disease Model Development

The recombinant receptor is instrumental in creating experimental models of inflammatory and autoimmune conditions. IL-1RI signaling is implicated in multiple disease contexts, including allergic inflammation, central nervous system pathology, cardiac dysfunction, rheumatoid arthritis, and cystic fibrosis. Using recombinant IL-1RI allows you to establish controlled experimental systems to test therapeutic hypotheses and validate potential interventions.

Therapeutic Target Validation

Since only approximately 5% of IL-1 receptors need to be occupied to induce cell activation, recombinant IL-1RI provides a tool to understand receptor occupancy dynamics and develop more effective therapeutic strategies. This is particularly relevant for designing receptor antagonists and blocking agents.

Practical Advantages

Standardization and Reproducibility

Recombinant proteins offer consistent quality and purity across experiments, eliminating batch-to-batch variability associated with native sources. This standardization is critical for obtaining reproducible results and comparing data across multiple studies.

Flexibility in Experimental Design

The recombinant format allows you to use the receptor in various applications, including cell culture assays, ELISA standards, and in vivo studies. You can select formulations with or without carrier proteins depending on your specific experimental requirements.

Cost-Effectiveness

Recombinant production typically provides a more economical source of high-quality protein compared to purification from native tissues, allowing for more extensive experimental work within budget constraints.

Recombinant Mouse IL-1 RI can be used as a standard for quantification or calibration in ELISA assays, but only if the ELISA is specifically designed to detect IL-1 RI and the recombinant protein is validated for this purpose.

Key considerations:

• ELISA Standard Requirements: For quantitative ELISA, the standard must be the same analyte as the target detected by the assay antibodies, and its form (e.g., recombinant, natural) must be validated to produce a parallel dose-response curve to the natural protein in biological samples.
• IL-1 RI vs. IL-1β/IL-1α: Most commercial ELISA kits for IL-1 family proteins are designed to detect cytokines such as IL-1β or IL-1α, not the receptor (IL-1 RI). Using IL-1 RI as a standard in an IL-1β or IL-1α ELISA would not be appropriate, as the antibodies in those kits are specific for the cytokine, not the receptor.
• IL-1 RI-Specific ELISA: If your assay is designed to detect mouse IL-1 RI (the receptor), and the recombinant protein matches the native sequence and conformation, it can be used as a standard. Some ELISA kits and DuoSet development kits are specifically validated for quantifying mouse IL-1 RI using recombinant IL-1 RI as the standard. Always confirm that the recombinant standard is compatible with your assay format and antibody pair.
• Validation: The recombinant standard should be validated to ensure that its dose-response curve is parallel to that of the native protein in your sample matrix, confirming accurate quantification. This is typically established by the ELISA kit manufacturer or through in-house validation.

Summary Table:

Best Practices:

• Always use the standard recommended by your ELISA kit or validate the recombinant protein as a standard in your specific assay.
• Check the product datasheet or technical documentation for validation data regarding use as a standard in ELISA.
• Ensure the recombinant protein is of high purity, correctly folded, and free of interfering substances.

If you are running an ELISA for IL-1 RI quantification, and your recombinant Mouse IL-1 RI is validated for this use, it is appropriate as a standard. For cytokine (IL-1β/IL-1α) quantification, use the corresponding recombinant cytokine standard.

Recombinant Mouse IL-1 RI has been validated for several key applications in published research, primarily in studies of cytokine signaling, inflammation, and immune response.

Validated Applications:

• Cytokine Bioassays: Recombinant Mouse IL-1 RI is used to study IL-1 signaling pathways, including the binding and response to IL-1α and IL-1β in cell-based assays.
• ELISA Standard: It serves as a standard or control in ELISA development for quantifying IL-1 or related cytokines in biological samples.
• Western Blotting: The protein is used as a positive control or for detection of IL-1 RI expression in tissue homogenates and cell lysates.
• Neutralization Assays: Recombinant Mouse IL-1 RI is employed to assess the neutralizing activity of antibodies or antagonists targeting IL-1 signaling, often in proliferation assays using T cell lines.
• Mass Cytometry (CyTOF): It is validated for use in high-dimensional single-cell analysis to study immune cell populations and cytokine receptor expression.
• Spatial Biology: Used in advanced imaging and tissue profiling to map IL-1 RI distribution and function in situ.
• Vaccine Development: Recombinant Mouse IL-1 RI is applied in preclinical studies to evaluate immune modulation and adjuvant effects.

Supporting Context:

• In genetic mouse models, recombinant IL-1 RI has been used to dissect cell-type-specific functions of IL-1 signaling, particularly in endothelial cells and leukocytes, demonstrating its utility in mechanistic studies of inflammation and CNS responses.
• It is also used in studies investigating the role of IL-1 RI in disease models such as myeloproliferative neoplasms, neuroinflammation, and autoimmune conditions.

Additional Notes:

• Recombinant Mouse IL-1 RI is typically used in in vitro and ex vivo assays, but also in in vivo models to study receptor function and therapeutic interventions.
• Validation for each application is supported by published research and product datasheets, confirming its biological activity and specificity in these contexts.

If you require protocol details or specific assay conditions, please specify the intended application.

To reconstitute and prepare Recombinant Mouse IL-1 RI protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
2. Reconstitute the protein in sterile, deionized water or a suitable buffer (such as 20 mM Tris, 150 mM NaCl, pH 8.0) to a final concentration of 0.1–1.0 mg/mL. Avoid concentrations below 100 μg/mL unless specifically required.
3. Gently mix by swirling or gentle pipetting. Avoid vigorous shaking or vortexing, as this can denature the protein or cause foaming.
4. Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
5. Aliquot the reconstituted protein into single-use portions to avoid repeated freeze-thaw cycles, which can degrade the protein.
6. Storage:
   • Store aliquots at 2–8 °C for up to 1 month under sterile conditions.
   • For longer-term storage, keep at –20 °C to –70 °C for up to 3 months.
7. For cell culture use, dilute the aliquoted stock to the desired working concentration using cell culture medium or a buffer containing a carrier protein (such as heat-inactivated FCS or tissue culture-grade BSA) to minimize adsorption and stabilize the protein.
8. Optional: For enhanced stability, you may add 5–50% glycerol to the reconstituted solution before freezing.

Key technical notes:

• Always use low endotoxin reagents and sterile technique to prevent contamination and ensure suitability for cell culture.
• If the protein is sensitive or you observe precipitation, consider using a buffer (e.g., Tris/NaCl) instead of water for reconstitution.
• Avoid repeated freeze-thaw cycles by aliquoting immediately after reconstitution.

Summary Table: Recombinant Mouse IL-1 RI Reconstitution

These steps will ensure the recombinant protein is properly prepared for reliable and reproducible cell culture experiments.