Recombinant Mouse IL-12 Receptor Beta 1 in Research Applications

Recombinant mouse IL-12 receptor beta 1 (IL-12Rβ1) serves as a critical tool for investigating IL-12 signaling pathways and immune responses. Here are the key reasons to incorporate this reagent into your research:

Mechanistic Studies of IL-12 Signaling

IL-12Rβ1 is an essential component of the IL-12 receptor complex, making recombinant versions invaluable for dissecting receptor-ligand interactions and downstream signaling cascades. By using the recombinant form, you can conduct controlled experiments to define functional binding sites and characterize how IL-12 engages with its receptor in a defined system. This allows you to map critical interaction domains and understand the molecular basis of IL-12-mediated immune activation.

Immunological Research Applications

The IL-12 receptor system plays a central role in T helper cell differentiation and immune responses. Recombinant IL-12Rβ1 enables you to:

• Study Th1 and Th17 differentiation: IL-12 signaling through IL-12Rβ1 drives distinct T cell lineage commitments, and recombinant receptor allows controlled investigation of these pathways
• Investigate signal transduction: The receptor activates JAK2 and TYK2 kinases, leading to STAT3 and STAT4 phosphorylation, which you can monitor in vitro using recombinant components
• Examine cancer immunotherapy mechanisms: Understanding IL-12Rβ1 function is relevant for developing improved cancer immunotherapies, as IL-12 demonstrates potent antitumor activity through enhanced NK cell and CD8+ T cell responses

Therapeutic Development and Optimization

Recombinant IL-12Rβ1 can support the development of next-generation IL-12-based therapeutics by enabling structure-function studies that inform rational design of improved cytokine variants or receptor-targeting strategies. This is particularly relevant given that systemic IL-12 administration has historically been limited by toxicity concerns, and understanding receptor biology may facilitate safer therapeutic approaches.

Recombinant Mouse IL-12 Rβ1 is a receptor subunit, not the cytokine itself, and is not suitable as a standard for quantification or calibration in ELISA assays designed to measure IL-12 or IL-12 p70. ELISA standards must match the analyte being measured—in this case, the cytokine IL-12 (typically the p70 heterodimer), not its receptor.

Key points:

• ELISA standards for IL-12 quantification are typically recombinant mouse IL-12 p70 protein, not the IL-12 receptor or its subunits.
• The IL-12 Rβ1 protein is a component of the IL-12 receptor complex and does not share antigenic or functional equivalence with IL-12 cytokine; thus, it will not be recognized by antibodies specific for IL-12 in standard ELISA kits.
• Using the receptor protein as a standard would result in no signal or inaccurate quantification, as the assay antibodies are not designed to detect IL-12 Rβ1.

Best practice:

• Use a recombinant mouse IL-12 p70 protein that matches the analyte for your ELISA standard curve.
• Confirm that the standard is validated for use in your specific ELISA format and is supplied or recommended by the kit manufacturer or validated in the literature.

If your goal is to quantify IL-12 cytokine, do not use IL-12 Rβ1 as a standard. Always match the standard to the analyte being measured for accurate and reliable ELISA results.

Recombinant Mouse IL-12 Rβ1 has been validated in published research primarily for applications involving the study of cytokine receptor interactions, immune cell signaling, and functional assays related to the IL-12 family of cytokines.

Key validated applications include:

• Binding and receptor-ligand interaction studies: Recombinant Mouse IL-12 Rβ1 is used to characterize the binding of IL-12 and related cytokines to their receptor complexes, enabling detailed analysis of receptor specificity and affinity.
• Cell signaling assays: It is employed to investigate downstream signaling events such as JAK2/TYK2 activation and STAT3/STAT4 phosphorylation upon ligand binding, which are central to understanding IL-12-mediated immune responses.
• Functional immunology assays: The protein is used in experiments to assess its role in T cell differentiation (e.g., Th1 and Th17), NK cell activation, and modulation of immune cell function in vitro and in vivo.
• Disease model studies: Recombinant Mouse IL-12 Rβ1 has been utilized in models of infection, cancer, and autoimmune disease to dissect the contribution of IL-12 signaling to disease progression and immune regulation.
• Evolutionary and comparative immunology: It serves as a tool for cross-species analyses of IL-12 family receptor evolution and function, as demonstrated in studies comparing receptor targets across multiple vertebrate species.

These applications are supported by published research focusing on the molecular mechanisms of IL-12 signaling, immune cell activation, and the therapeutic potential of modulating IL-12 pathways in disease contexts. The use of recombinant receptor proteins like IL-12 Rβ1 is essential for dissecting cytokine-receptor interactions and validating targets for immunomodulatory therapies.

To reconstitute and prepare Recombinant Mouse IL-12 Rβ1 protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS.

Essential context and protocol details:

• Reconstitution:
  Add sterile PBS directly to the lyophilized vial to achieve a final concentration of 100 μg/mL. Gently swirl or invert the vial to mix; avoid vigorous shaking to prevent protein denaturation.

• Dilution for cell culture:
  After reconstitution, further dilute the stock solution to your desired working concentration using sterile cell culture medium or PBS. If your application is sensitive to protein adsorption or loss, consider adding a carrier protein such as 0.1% BSA to the dilution buffer, unless your experiment requires carrier-free conditions.

• Sterility:
  Ensure all solutions and materials are sterile to prevent contamination of cell cultures.

• Storage:

  • After reconstitution, the protein is stable for up to 1 month at 2–8 °C under sterile conditions.
  • For longer-term storage (up to 3 months), aliquot and store at –20 °C to –70 °C to avoid repeated freeze-thaw cycles.
  • Use a manual defrost freezer and avoid repeated freeze-thaw cycles to maintain protein integrity.

• General best practices:

  • Thaw aliquots on ice and mix gently before use.
  • Discard any unused reconstituted protein that has been stored at 2–8 °C for more than 1 month or has undergone multiple freeze-thaw cycles.

Summary Table: Recombinant Mouse IL-12 Rβ1 Reconstitution

This protocol ensures the protein remains stable and bioactive for cell culture applications. If your experimental design requires carrier-free conditions, confirm that no stabilizers or carrier proteins are present in your reconstitution or dilution buffers.