Recombinant Human IL-1Rrp2

Recombinant human IL-1Rrp2 (also known as IL-1R6) serves as a valuable research tool for investigating IL-1 family cytokine signaling and immune regulation across multiple disease contexts.

Key Research Applications

Immune Checkpoint and Inflammatory Signaling Studies

IL-1Rrp2 functions as a stimulatory immune checkpoint in various cell types and tissues. This receptor mediates activation of the transcription factor NF-κB in response to IL-1 family ligands, particularly IL-1F6, IL-1F8, and IL-1F9. Using recombinant IL-1Rrp2 allows you to investigate how these novel IL-1 cytokines activate downstream signaling cascades, including MAP kinase pathways and transcription factor activation.

Disease Modeling and Therapeutic Development

The receptor is highly expressed in skin and multiple skin-related cell types, making it particularly relevant for studying skin tumorigenesis and inflammatory skin diseases. IL-1Rrp2 expression patterns also appear significant in brain cells and glial cells, enabling investigation of neuroinflammatory processes. Recombinant IL-1Rrp2 can be used to model disease mechanisms in conditions where IL-1 family cytokines play pathogenic roles, including arthritis, endometriosis, organ transplantation complications, and autoimmune conditions.

Signal Transduction Research

The IL-1Rrp2/JNK pathway represents an important signaling axis for inflammatory cytokine expression. Recombinant IL-1Rrp2 enables detailed mechanistic studies of how IL-1 family ligands activate this pathway and downstream inflammatory responses, facilitating the development of targeted therapeutic interventions.

Comparative Receptor Studies

Since IL-1Rrp2 works in conjunction with IL-1RAcP as a co-receptor for certain IL-1 family members, recombinant IL-1Rrp2 allows investigation of receptor heterodimerization, ligand specificity, and differential signaling outcomes compared to other IL-1 receptor family members.

You can use recombinant Human IL-1Rrp2 as a standard for quantification or calibration in ELISA assays, provided that the recombinant protein is of high purity, its concentration is accurately determined, and it is compatible with the antibodies and detection system used in your assay.

For ELISA quantification:

• Recombinant proteins are commonly used as standards in ELISA assays when purified native protein is unavailable.
• The standard curve should be prepared using serial dilutions of the recombinant IL-1Rrp2 in the same buffer as your samples to ensure matrix compatibility.
• Accurate quantification requires that the recombinant standard is well-characterized, ideally with concentration confirmed by an independent method such as absorbance at 280 nm or HPLC.
• The standard must be reconstituted and diluted according to best practices, avoiding conditions that could denature or aggregate the protein.

Critical considerations:

• The ELISA antibodies must recognize both the recombinant and native forms of IL-1Rrp2 with similar affinity. If the recombinant protein differs in glycosylation or folding from the native protein, this could affect assay accuracy.
• The standard curve should cover the expected concentration range of your samples, typically from low pg/mL to ng/mL, depending on assay sensitivity.
• Always run a standard curve with each assay to ensure reliable quantification.

Best practices:

• Validate the recombinant standard in your assay by spiking known amounts into sample matrix and assessing recovery and linearity.
• Document the source, lot, and characterization data of the recombinant protein for reproducibility.

In summary, recombinant Human IL-1Rrp2 is suitable as an ELISA standard if it is pure, accurately quantified, and compatible with your assay reagents and conditions.

Recombinant Human IL-1Rrp2 (IL-1R6/IL-36R) has been validated for several key applications in published research, primarily focusing on its role in immunological and inflammatory signaling pathways. The main applications include:

1. Protein-Protein Interaction Studies

   • IL-1Rrp2 has been used to investigate its interaction with ligands such as IL-36β and IL-38. For example, functional ELISA assays have demonstrated its binding ability to IL-36β, confirming its role as a receptor for IL-36 family cytokines (Assay Genie, Fisher Scientific, R&D Systems).

2. Signal Transduction and Pathway Activation

   • Studies have shown that IL-1Rrp2 is essential for IL-38-induced activation of the JNK and AP-1 pathways in transfected HEK293T cells. This demonstrates its involvement in downstream inflammatory signaling (PMC9171418).

3. Inflammatory Cytokine Regulation

   • IL-1Rrp2 has been validated for its role in stimulating the expression of inflammatory cytokines in tumor cells. Knockdown experiments using siRNA against IL-1Rrp2 have shown reduced cytokine production upon IL-38 stimulation, highlighting its function as a stimulatory immune checkpoint in skin cancer (PMC9171418).

4. Therapeutic Target Validation

   • The receptor has been used in studies evaluating the effects of IL-36 receptor antagonists, such as in models of liver damage and inflammatory skin diseases. These studies demonstrate its relevance as a therapeutic target for inflammatory and autoimmune conditions (Nature Scientific Reports).

5. Expression and Functional Analysis in Cell Lines

   • IL-1Rrp2 expression has been analyzed in various cell types, including skin cancer cell lines (A431) and glial cells, confirming its tissue-specific expression and functional relevance in different biological contexts (PMC9171418, Manchester Research Portal).

6. Neutralization and Blocking Assays

   • Antibodies against IL-1Rrp2 have been used to neutralize IL-36γ-induced IL-8 secretion in epithelial carcinoma cell lines, validating its role in mediating inflammatory responses (R&D Systems).

In summary, Recombinant Human IL-1Rrp2 has been validated for applications in protein interaction studies, signal transduction, inflammatory cytokine regulation, therapeutic target validation, and functional analysis in various cell types, making it a valuable tool for immunological and inflammatory disease research.

To reconstitute and prepare Recombinant Human IL-1Rrp2 (IL-1R6/IL1RL2) protein for cell culture experiments, follow these steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute the protein in sterile distilled water to a final concentration of 0.1–0.5 mg/mL.
• Avoid vortexing or vigorous pipetting; gently mix by swirling or inverting to prevent protein denaturation.
• For long-term storage, add a carrier protein or stabilizer (e.g., 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose) to the reconstituted solution, then aliquot and store at ≤ –20°C to minimize freeze-thaw cycles.
• After reconstitution, the solution is stable at –20°C for up to 3 months and at 2–8°C for up to 1 week; avoid repeated freeze/thaw cycles.
• For cell culture use, dilute the reconstituted stock into your culture medium or buffer immediately before use, ensuring sterility throughout the process.

Additional notes for best practice:

• Always consult the specific product datasheet for recommended diluents and concentrations, as some formulations may require PBS instead of water.
• If the protein is fused to an Fc tag or other domains, confirm compatibility with your assay or cell system.
• If using for functional assays, verify bioactivity post-reconstitution if possible.

Summary protocol:

1. Centrifuge vial, open in sterile conditions.
2. Add sterile distilled water (or PBS if specified) to achieve 0.1–0.5 mg/mL.
3. Gently mix; do not vortex.
4. Add carrier protein/stabilizer for storage if needed.
5. Aliquot and store at ≤ –20°C.
6. Dilute into cell culture medium for experiments.

This protocol ensures optimal solubility, stability, and activity of recombinant IL-1Rrp2 for cell culture applications.