Recombinant Human IL-10 Rα

Recombinant Human IL-10 Rα is used in research applications to study and manipulate the IL-10 signaling pathway, which is central to immune regulation, inflammation control, and disease modeling.

Key scientific applications and rationale:

• Mechanistic Studies: IL-10 Rα is the primary receptor subunit for IL-10, mediating its anti-inflammatory effects through activation of the JAK-STAT signaling pathway, particularly STAT3. Recombinant IL-10 Rα enables detailed investigation of receptor-ligand interactions, downstream signaling events, and transcriptional responses in various cell types.

• Functional Assays: Recombinant IL-10 Rα can be used to assess the biological activity of IL-10 or its variants, such as measuring inhibition of IL-10-dependent cell proliferation (e.g., MC/9-2 mouse mast cells). This is essential for validating the potency and specificity of IL-10 or engineered cytokines in vitro.

• Disease Modeling: Deficiencies or mutations in IL-10 or IL-10Rα are linked to inflammatory diseases such as colitis and early-onset inflammatory bowel disease. Recombinant IL-10 Rα allows researchers to model these conditions, study genetic susceptibility, and test therapeutic interventions targeting the IL-10 pathway.

• Therapeutic Development: IL-10 and its receptor are attractive targets for drug development in autoimmune diseases, cancer, and tissue damage. Recombinant IL-10 Rα is used in screening and optimizing biologics, small molecules, or engineered cytokines that modulate IL-10 signaling.

• Cellular and Molecular Biology: Recombinant IL-10 Rα is valuable for receptor binding studies, affinity measurements, and structural biology, helping elucidate how IL-10 variants or antagonists interact with the receptor and influence immune cell function.

Best practices:

• Use recombinant IL-10 Rα in controlled in vitro assays to quantify IL-10 activity and receptor engagement.
• Employ it in cell-based models to dissect signaling pathways and gene expression profiles downstream of IL-10.
• Integrate recombinant receptor studies with genetic or pharmacological approaches to validate findings in disease-relevant contexts.

Summary:
Using recombinant human IL-10 Rα in research provides a precise tool to dissect IL-10-mediated immune regulation, validate therapeutic candidates, and model disease mechanisms involving the IL-10 pathway.

No, you cannot use recombinant human IL-10 Rα as a standard for quantification or calibration in IL-10 ELISA assays. IL-10 Rα is a receptor protein, not the cytokine itself, and serves a fundamentally different biological role than IL-10.

Why IL-10 Rα Is Unsuitable as an ELISA Standard

Structural and Functional Differences

IL-10 Rα is the alpha subunit of the IL-10 receptor complex and functions as a binding partner for IL-10, not as the measurand. ELISA assays designed to quantify IL-10 use antibodies specifically raised against the IL-10 cytokine protein itself. These antibodies will not recognize IL-10 Rα with the same affinity or specificity, if at all.

Cross-Reactivity Testing

Specificity studies on IL-10 ELISA kits have explicitly evaluated IL-10 Rα as a potential interferent. Testing shows that IL-10 Rα does not cross-react with the assay components designed to detect IL-10. This lack of cross-reactivity confirms that IL-10 Rα will not generate a measurable signal in an IL-10-specific sandwich ELISA.

Proper ELISA Calibration Standards

IL-10 ELISA assays are calibrated using recombinant human IL-10 protein itself, typically expressed in insect cells or mammalian expression systems. These standards are highly purified (≥95% purity) and are traceable to international reference materials such as the NIBSC/WHO reference reagent. The standard curves are generated using serial dilutions of recombinant IL-10, typically ranging from 2-500 pg/mL depending on the assay platform.

For accurate quantification of IL-10 in your samples, use only recombinant human IL-10 protein as your calibration standard, ensuring it is from a validated source and appropriate for your specific ELISA platform.

Recombinant Human IL-10 Rα (Interleukin-10 Receptor alpha) has been validated in published research for several key applications, primarily focused on its role in immunological assays and mechanistic studies of IL-10 signaling.

Validated Applications in Published Research:

• Bioassays: Recombinant human IL-10 and its receptor components, including IL-10 Rα, are widely used in cell-based bioassays to study anti-inflammatory signaling, cytokine response, and immune cell modulation. These assays often involve primary human monocytes, T cells, or other immune cell types to assess downstream effects such as STAT1/STAT3 phosphorylation, gene expression changes, and cytokine secretion.

• ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant IL-10 Rα is used as a standard or capture/detection reagent in ELISA protocols for quantifying IL-10 or monitoring IL-10/IL-10R interactions in biological samples.

• Blocking/Neutralization Studies: Recombinant IL-10 Rα is employed in functional assays to block or neutralize IL-10 signaling, enabling researchers to dissect the specific contributions of IL-10/IL-10R interactions in immune regulation and inflammation.

• Receptor-Ligand Binding Studies: Recombinant IL-10 Rα is used in binding assays (e.g., surface plasmon resonance, yeast surface display) to characterize the affinity and kinetics of IL-10 or engineered IL-10 variants for their receptor, and to study receptor complex formation and stoichiometry.

• Therapeutic Mechanism Studies: Recombinant IL-10 Rα is utilized in preclinical models to investigate the therapeutic potential of IL-10-based interventions, including engineered IL-10 variants with altered receptor affinity, and to evaluate anti-inflammatory efficacy in vitro and in vivo.

Supporting Details and Examples:

• In studies engineering IL-10 variants, recombinant IL-10 Rα was used to assess enhanced receptor binding and downstream signaling, such as STAT1/3 activation in human monocytes and CD8+ T cells.
• ELISA protocols have used recombinant IL-10 Rα as a standard for quantifying IL-10 in patient samples, particularly in autoimmune disease research.
• Blocking assays with recombinant IL-10 Rα have helped delineate the specific role of IL-10 signaling in suppressing inflammatory cytokine production and modulating immune cell phenotypes.
• Recombinant IL-10 Rα has been instrumental in studies of autoimmune diseases, tissue injury, and cancer, where modulation of IL-10 signaling is a therapeutic target.

Summary Table: Applications of Recombinant Human IL-10 Rα

These applications are supported by a broad range of immunology and cytokine research, particularly in the context of inflammation, autoimmunity, and therapeutic development.

To reconstitute and prepare Recombinant Human IL-10 Rα protein for cell culture experiments, dissolve the lyophilized protein at 100 μg/mL in sterile PBS (phosphate-buffered saline). For optimal stability and activity, it is recommended to include at least 0.1% human or bovine serum albumin (HSA/BSA) in the PBS when preparing working solutions, especially for cell culture applications.

Step-by-step protocol:

• Briefly centrifuge the vial to collect the lyophilized protein at the bottom before opening.
• Add sterile PBS (pH 7.2–7.4) to achieve a final concentration of 100 μg/mL.
• If the protein formulation does not already contain carrier protein, supplement the PBS with 0.1% HSA or BSA to minimize adsorption and loss.
• Gently swirl or tap the vial to mix; avoid vigorous pipetting or vortexing to prevent protein denaturation.
• Allow the protein to fully dissolve (typically 5–10 minutes at room temperature).
• Once reconstituted, aliquot the solution to avoid repeated freeze-thaw cycles and store at –20°C or below.
• For cell culture, further dilute the stock solution in your culture medium immediately before use.

Additional notes:

• Always use sterile technique to prevent contamination.
• Avoid repeated freeze-thaw cycles, as these can reduce protein activity.
• If the protein is supplied with BSA or another carrier, direct reconstitution in PBS is sufficient; otherwise, adding carrier protein is strongly recommended for cell-based assays.
• For functional assays, confirm the final working concentration based on experimental requirements (e.g., dose-response studies).

This protocol ensures maximal stability and bioactivity of recombinant IL-10 Rα for cell culture experiments.