IL10RB Antibody

What is IL10RB and what is its function in the immune system?

IL10RB (interleukin 10 receptor subunit beta) is a transmembrane protein that functions as a subunit of the interleukin-10 receptor complex. It is also known by several other names including CDW210B, CRF2-4, CRFB4, D21S58, and IL-10 receptor subunit beta. Structurally, the protein has a molecular mass of approximately 37 kilodaltons .

The IL10RB subunit forms heterodimers with other receptor subunits to create functional receptors for several cytokines, primarily IL-10. This receptor complex plays a critical role in immune tolerance and mucosal homeostasis. IL-10 signaling through its receptor is essential for regulating inflammatory responses and maintaining intestinal immune balance. Studies in mouse models have demonstrated that IL10RB deficiency leads to spontaneous colitis, highlighting its importance in preventing excessive inflammation in the intestine .

What applications are IL10RB antibodies commonly used for?

IL10RB antibodies are utilized across multiple research applications, including:

• Western Blot (WB): For detecting IL10RB protein in cell or tissue lysates, with an observed molecular weight of 42-45 kDa

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative detection of IL10RB

• Flow Cytometry (FCM): For analyzing IL10RB expression on cell surfaces

• Immunocytochemistry (ICC) and Immunofluorescence (IF): For visualizing IL10RB in cells

• Immunohistochemistry on frozen (IHC-fr) or paraffin-embedded (IHC-p) tissues: For examining IL10RB expression in tissue sections

• Neutralization assays (Neut): For studying functional aspects of IL10RB

The choice of application should be guided by the specific research question and experimental design.

What species reactivity should I consider when selecting an IL10RB antibody?

When selecting an IL10RB antibody, it's important to consider the species used in your experiments. Commercial antibodies are available with reactivity to human, mouse, and rat IL10RB . Based on gene sequence homology, antibodies may also detect IL10RB orthologs in other species such as canine, porcine, and monkey models, though this should be validated experimentally .

For cross-species studies, it's advisable to select antibodies that have been validated for reactivity with IL10RB from all relevant species or to perform validation studies before proceeding with full experiments.

How should I optimize Western blot protocols for IL10RB detection?

For optimal Western blot detection of IL10RB:

• Sample preparation: IL10RB has been successfully detected in mouse skeletal muscle tissue and Jurkat cells . For human samples, consider using immune cells, particularly macrophages or lymphocytes where IL10RB is highly expressed.

• Expected molecular weight: While the theoretical molecular weight of IL10RB is approximately 37 kDa , the observed molecular weight by Western blot is typically 42-45 kDa . This discrepancy may be due to post-translational modifications such as glycosylation.

• Antibody dilution: Start with a dilution range of 1:200-1:2000 for Western blot applications . Optimization may be required for your specific experimental conditions.

• Controls: Include positive controls such as Jurkat cell lysates or mouse skeletal muscle tissue . For negative controls, consider using tissues from IL10RB knockout models if available, or samples where IL10RB expression has been silenced.

• Validation: Confirm specificity using multiple antibodies targeting different epitopes of IL10RB to ensure consistent results.

What are the key considerations for immunohistochemical detection of IL10RB?

When performing immunohistochemistry for IL10RB:

• Tissue fixation and processing: Both frozen (IHC-fr) and paraffin-embedded (IHC-p) tissues can be used for IL10RB detection . Optimize antigen retrieval methods for paraffin sections, as excessive fixation may mask the epitope.

• Antibody selection: Choose antibodies specifically validated for IHC applications. Polyclonal antibodies may offer better sensitivity for IHC, especially for proteins with lower expression levels.

• Signal amplification: Consider using polymer-based detection systems or tyramide signal amplification to enhance sensitivity, particularly in tissues with lower IL10RB expression.

• Co-localization studies: Combine IL10RB staining with markers for specific immune cell populations (such as macrophages or T cells) to identify cell types expressing IL10RB in tissue contexts.

• Interpretation: IL10RB expression may vary across different tissue compartments and disease states. In inflammatory conditions, expect potentially altered expression patterns compared to normal tissues.

How can IL10RB antibodies be used to study inflammatory bowel disease mechanisms?

IL10RB deficiency is associated with early-onset inflammatory bowel disease (IBD) . Researchers investigating IBD mechanisms can utilize IL10RB antibodies in several advanced applications:

• Macrophage polarization analysis: IL10RB signaling significantly impacts macrophage differentiation into pro-inflammatory versus anti-inflammatory phenotypes . Using flow cytometry with IL10RB antibodies alongside markers for M1 (pro-inflammatory) and M2 (anti-inflammatory) macrophages can reveal how IL10RB expression correlates with functional polarization in patient samples.

• T-regulatory cell function: IL10RB signaling on innate immune cells is crucial for the suppressive function of regulatory T cells . Implement co-culture experiments with macrophages and Treg cells, using IL10RB antibodies to monitor receptor expression and neutralizing antibodies to block signaling.

• Tissue-specific expression patterns: Compare IL10RB expression in inflamed versus non-inflamed intestinal tissues from IBD patients using immunohistochemistry to identify spatial patterns that correlate with disease activity.

• Genetic correlation studies: For patients with IL10RB mutations, combine genetic analysis with protein expression studies using specific antibodies that can detect mutant forms to establish genotype-phenotype correlations.

What role does IL10RB play in viral infections and how can antibodies help investigate this?

IL10RB has been identified as a key regulator of COVID-19 host susceptibility, with higher expression associated with worse disease outcomes . Researchers can use IL10RB antibodies to investigate viral infection mechanisms through:

• Expression analysis during infection: Monitor changes in IL10RB expression levels in response to viral infection using Western blot or flow cytometry with IL10RB antibodies.

• Viral load correlation: Establish correlations between IL10RB expression levels (quantified by antibody-based methods) and viral load in experimental infection models.

• Signaling pathway investigation: Use phospho-specific antibodies against downstream signaling molecules in the IL10R pathway alongside IL10RB antibodies to track how receptor engagement affects intracellular signaling during viral infection.

• Neutralization studies: Employ neutralizing antibodies against IL10RB to block receptor function and assess the impact on viral replication and immune response activation in cell culture models.

• Single-cell analysis: Combine IL10RB antibodies with single-cell technologies to identify which specific immune cell populations alter IL10RB expression during viral infection.

How can I validate the specificity of IL10RB antibodies?

Ensuring antibody specificity is crucial for reliable results. Consider these validation approaches:

• Genetic controls: Test antibodies on samples from IL10RB knockout models or cells where IL10RB has been silenced via siRNA/CRISPR techniques.

• Peptide competition: Pre-incubate the antibody with excess immunizing peptide before application to samples. A specific antibody should show diminished or absent signal.

• Multiple antibody approach: Use different antibodies targeting distinct epitopes of IL10RB and compare detection patterns.

• Correlation with mRNA expression: Compare protein detection levels with mRNA expression data from RT-PCR or RNA-seq.

• Mass spectrometry validation: For critical applications, consider immunoprecipitation followed by mass spectrometry to confirm that the protein being detected is indeed IL10RB.

What are common issues with IL10RB detection and how can they be resolved?

Several challenges may arise when working with IL10RB antibodies:

• Inconsistent molecular weight: The observed molecular weight (42-45 kDa) differs from the theoretical weight (37 kDa) due to post-translational modifications. Use positive controls with known IL10RB expression to establish the expected band pattern for your system.

• Low expression levels: In some cell types or tissues, IL10RB expression may be below detection threshold. Consider:

  • Signal amplification methods

  • Enrichment of membrane fractions for Western blot

  • Longer exposure times (while monitoring background)

  • More sensitive detection substrates

• Cross-reactivity: Some antibodies may cross-react with related receptors. Validate specificity using the methods described above and analyze the epitope sequence for homology with other proteins.

• Fixation artifacts in IHC: Optimize fixation protocols and antigen retrieval methods. Compare results from frozen and paraffin-embedded tissues to identify potential fixation-related issues.

How can IL10RB antibodies be used to study the relationship between IL10RB deficiency and early-onset IBD?

IL10RB deficiency is associated with very early-onset IBD, typically manifesting within the first months of life . Researchers studying this relationship can:

• Patient sample analysis: Use IL10RB antibodies to assess protein expression in patient-derived samples, correlating expression levels with disease severity and specific mutations.

• Functional studies: Implement IL10RB antibodies in assays measuring cytokine production, cell proliferation, and apoptosis to understand how IL10RB deficiency affects cellular functions.

• Therapeutic monitoring: For patients receiving experimental therapies, monitor IL10RB expression and downstream signaling responses using antibody-based detection methods.

• Mechanistic investigation: Compare IL10RA and IL10RB-deficient models to distinguish shared versus unique pathways, as the clinical manifestations differ between these two deficiencies .

• Biomarker development: Investigate whether soluble forms of IL10RB could serve as biomarkers for disease progression or treatment response using capture antibodies in ELISA formats.

What experimental approaches can clarify the distinct roles of IL10RB in different immune cell populations?

IL10RB functions may vary across immune cell types. To investigate these distinctions:

• Cell-specific knockout models: Study the consequences of IL10RB deletion in specific cell lineages (macrophages, dendritic cells, T cells) using conditional knockout models and monitor changes with IL10RB antibodies.

• Single-cell protein analysis: Combine IL10RB antibodies with markers for specific immune cell subsets in flow cytometry or mass cytometry to analyze expression patterns at the single-cell level.

• Bone marrow chimeras: Create mixed bone marrow chimeras with wild-type and IL10RB-deficient cells to assess cell-intrinsic versus cell-extrinsic effects, using IL10RB antibodies to track reconstitution.

• Ex vivo functional assays: Isolate specific immune cell populations and assess their response to IL10 and other IL10RB-binding cytokines, correlating functional outcomes with receptor expression levels measured by antibody-based methods.

• Spatial transcriptomics integration: Combine IL10RB immunohistochemistry with spatial transcriptomics to correlate protein expression with transcriptional programs in distinct microanatomical niches.