Recombinant Rat IL-1Rrp2 (also known as IL-1 receptor-like 2 or IL-1R6) is valuable in research applications focused on understanding and modulating the IL-1 family cytokine signaling, particularly for studies involving inflammation, immune regulation, and neurobiology.

Key reasons to use recombinant Rat IL-1Rrp2 in research:

• Specificity for IL-1 Family Ligands: IL-1Rrp2 is the receptor for newer IL-1 family cytokines such as IL-1F9 (also known as IL-36γ), which are implicated in inflammatory and autoimmune processes. Using recombinant IL-1Rrp2 allows you to study ligand-receptor interactions, signaling pathways, and downstream effects in a controlled system.

• Cellular Targeting and Expression: High expression of IL-1Rrp2 mRNA has been detected in glial cells, suggesting its importance in neuroinflammation and brain cell signaling. Recombinant protein enables in vitro and in vivo experiments to dissect receptor function in specific cell types.

• Functional Studies: Recombinant IL-1Rrp2 can be used to:

  • Investigate receptor-ligand binding kinetics.
  • Assess downstream signaling events (e.g., activation of nuclear factor pathways).
  • Screen for antagonists or agonists that modulate IL-1Rrp2 activity.
  • Model disease states where IL-1Rrp2-mediated signaling is dysregulated.

• Protein Engineering and Therapeutic Development: The recombinant form, especially as an Fc chimera, offers enhanced stability and shelf-life, facilitating biochemical assays, structural studies, and therapeutic screening.

• Inflammation and Immune Modulation: IL-1 family receptors, including IL-1Rrp2, play roles in acute and chronic inflammation, autoimmune diseases, and tissue homeostasis. Recombinant receptor proteins are essential for mechanistic studies and for developing targeted interventions.

• Neurobiology Applications: Since IL-1Rrp2 is expressed in brain cells and interacts with IL-1F9, it is particularly relevant for research into neuroinflammatory and neurodegenerative conditions.

Best Practices:

• Use recombinant IL-1Rrp2 in cell-based assays, binding studies, and animal models to elucidate its biological functions.
• Employ carrier proteins or Fc chimeras to improve protein stability and experimental reproducibility.
• Store recombinant proteins in polypropylene tubes and at recommended conditions to maintain activity.

In summary, recombinant Rat IL-1Rrp2 is a critical tool for dissecting IL-1 family signaling, understanding receptor biology in inflammation and neurobiology, and developing targeted modulators for therapeutic research.

Yes, Recombinant Rat IL-1Rrp2 can be used as a standard for quantification or calibration in ELISA assays, provided it is of sufficient purity and its concentration is accurately determined.

For ELISA quantification, the standard should be a purified recombinant protein that matches the analyte being measured. Recombinant proteins are commonly used as standards in ELISA protocols, as they allow for the generation of a standard curve to quantify unknown samples. The recombinant Rat IL-1Rrp2/IL-1R6 Fc Chimera Protein is specifically recommended for use as an ELISA standard, especially when formulated with BSA to enhance stability and solubility.

Best practices for using recombinant proteins as ELISA standards:

• Confirm the identity and purity of the recombinant protein, ideally by SDS-PAGE and mass spectrometry.
• Accurately determine the protein concentration using a reliable method (e.g., UV absorbance at 280 nm, BCA assay).
• Prepare the standard curve by serially diluting the recombinant protein in the same buffer as your samples to minimize matrix effects.
• Ensure the ELISA antibodies are specific for Rat IL-1Rrp2 and recognize the recombinant form used as the standard.

Additional considerations:

• If your ELISA kit is designed for Rat IL-1Rrp2/IL-1R6, confirm that the recombinant standard is compatible with the kit’s antibody pair and detection system.
• Follow manufacturer or published protocols for reconstitution and handling of the recombinant protein, as improper preparation can affect quantification accuracy.
• Validate the standard curve by comparing it to known concentrations or by spike-and-recovery experiments to ensure reliable quantification.

In summary, recombinant Rat IL-1Rrp2 is suitable as an ELISA standard if it is properly prepared and validated for your assay system.

Applications of Recombinant IL-1Rrp2 in Published Research

Recombinant IL-1Rrp2 (also designated IL-1R6 or IL-1RL2) has been validated for several important research applications based on published studies:

Receptor Characterization and Signaling Studies

Recombinant IL-1Rrp2 has been utilized to characterize receptor-ligand interactions and downstream signaling pathways. In particular, recombinant IL-1Rrp2 has been employed in transfection studies to investigate IL-38-induced signaling, where cells transfected with full-length IL-1Rrp2 demonstrated enhanced AP-1 activation upon IL-38 stimulation compared to control vectors. This application has been critical for elucidating the IL-1Rrp2/JNK signaling axis in inflammatory responses.

Ligand Binding and Functional Analysis

The recombinant protein has been validated for investigating ligand-receptor interactions with IL-1 family cytokines. Research has demonstrated that IL-1Rrp2 serves as a functional receptor for IL-1F9 (IL-1H1), with high expression of IL-1Rrp2 mRNA detected in glial cells, establishing this receptor as a target for novel IL-1 family ligands.

Standard Analytical Applications

Recombinant IL-1Rrp2 is commonly employed in standard immunological assays. The protein has been validated for use as an ELISA standard and in Western blot applications. Additionally, antibodies targeting IL-1Rrp2 have been validated for multiple applications including ELISA, flow cytometry, Western blot, immunofluorescence, and immunohistochemistry.

Cell Culture and Tissue Culture Applications

The recombinant protein is recommended for use in cell and tissue culture systems, particularly when formulated with carrier proteins such as BSA for maintaining protein stability and preventing non-specific adsorption.

To reconstitute and prepare Recombinant Rat IL-1Rrp2 protein for cell culture experiments, dissolve the lyophilized protein in sterile phosphate-buffered saline (PBS) at a concentration of 0.1–0.2 mg/mL, using gentle handling to preserve protein integrity.

Step-by-step protocol:

• Equilibrate the lyophilized vial and PBS to room temperature before opening.
• Centrifuge the vial briefly (20–30 seconds in a microcentrifuge) to collect all powder at the bottom.
• Add sterile PBS slowly to the vial to reach the desired concentration (commonly 0.1–0.2 mg/mL for IL-1Rrp2). Avoid exceeding 1 mg/mL to prevent solubility issues.
• Gently mix by swirling or pipetting up and down. Do not vortex, as vigorous mixing can denature the protein.
• Allow to dissolve at room temperature for 15–30 minutes with gentle agitation. If undissolved material remains, extend gentle mixing up to 2 hours.
• Aliquot the reconstituted protein into single-use volumes (≥20 μL) to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at –80 °C for long-term storage. Avoid repeated freeze/thaw cycles.

Additional notes for cell culture use:

• For further dilution into cell culture medium, use sterile PBS or medium containing a carrier protein (e.g., 0.1% BSA, FBS, or HSA) to minimize adsorption and stabilize the protein.
• Confirm the final working concentration based on your experimental requirements and titrate as needed.
• If the protein is a fusion (e.g., Fc chimera), ensure compatibility with your assay system.

Summary of best practices:

• Use sterile technique throughout.
• Avoid vigorous mixing and repeated freeze/thaw.
• Use carrier proteins for dilution to working concentrations.

These steps will help maintain the biological activity and stability of recombinant rat IL-1Rrp2 for cell-based assays.