IL13RA1 Antibody, FITC conjugated

What is IL13RA1 and what cellular functions does it mediate?

IL13RA1 (Interleukin-13 receptor subunit alpha-1) is a receptor chain that binds with low affinity to interleukin-13 (IL13). When paired with IL4RA, it forms a functional receptor complex for IL13. Additionally, IL13RA1 serves as an alternate accessory protein to the common cytokine receptor gamma chain for interleukin-4 (IL4) signaling, though it cannot replace the function of IL2RG in enhancing interleukin-2 binding activity .

From a functional perspective, IL13RA1 plays critical roles in multiple physiological systems. Recent research has identified that IL13RA1 regulates myocardial homeostasis and cardiac function, with IL13ra1-deficient mice developing severe myocardial dysfunction and dyssynchrony . Beyond cardiac function, IL13RA1 is instrumental in mediating allergic airway responses, regulating inflammation, and participating in fibrotic processes in various tissues .

What are the key characteristics of IL13RA1 Antibody, FITC conjugated?

IL13RA1 Antibody, FITC conjugated is characterized by the following properties:

The antibody is supplied as a FITC-conjugated preparation, making it directly applicable for flow cytometry and immunofluorescence techniques without requiring secondary antibody labeling .

What are the validated applications for IL13RA1 Antibody, FITC conjugated in research settings?

The IL13RA1 Antibody, FITC conjugated has been validated for multiple research applications, though specific applications may vary between commercial products. Based on the available information, the following applications have been validated:

• Flow Cytometry: The FITC conjugation makes this antibody particularly suitable for direct detection in flow cytometry. Research has demonstrated successful detection of IL-13RA1 in human granulocytes using anti-IL13RA1 antibodies followed by fluorophore-conjugated secondary antibodies .

• Immunohistochemistry (IHC): IL-13RA1 has been successfully detected in paraffin-embedded tissue sections, including human skin and heart tissues. Protocols typically involve antigen retrieval, primary antibody incubation (5-15 μg/mL), and visualization using appropriate detection systems .

• Western Blotting: Some anti-IL13RA1 antibodies have been validated for Western blot applications, detecting the target protein in various human tissue lysates, including lung and pancreas .

• Direct ELISA: Anti-IL13RA1 antibodies have demonstrated utility in direct ELISA systems for detecting recombinant and native IL13RA1 proteins .

When establishing a new experimental protocol, researchers should perform appropriate validation steps including positive and negative controls to ensure specificity and optimal working conditions for their particular experimental system.

What is the recommended protocol for detecting IL13RA1 in tissue sections using immunohistochemistry?

Based on published methodologies, the following protocol is recommended for detecting IL13RA1 in tissue sections using immunohistochemistry:

• Tissue Preparation:

  • Fix tissue samples in 4% formaldehyde/paraformaldehyde

  • Embed in paraffin and section to 5 μm thickness

• Antigen Retrieval:

  • Perform heat-induced epitope retrieval using a basic antigen retrieval solution

  • Follow manufacturer's recommendations for specific buffer composition and heating conditions

• Primary Antibody Application:

  • Block non-specific binding with appropriate blocking solution

  • Apply IL13RA1 primary antibody at 5-15 μg/mL (concentration may need optimization)

  • Incubate either overnight at 4°C or for 1 hour at room temperature

• Detection:

  • For non-conjugated primary antibodies: Apply appropriate HRP-polymer detection system

  • For FITC-conjugated antibodies: Direct visualization under fluorescence microscopy

  • For chromogenic detection: Use DAB substrate and counterstain with hematoxylin

One validated approach documented in research used goat anti-human IL-13R alpha 1 antibody at 5 μg/mL for 1 hour at room temperature, followed by incubation with anti-goat IgG HRP polymer antibody, visualized with DAB and counterstained with hematoxylin. This method successfully detected specific staining in the cytoplasm of myocardial cells .

How can IL13RA1 Antibody, FITC conjugated be utilized to investigate cardiac pathophysiology?

Recent research has revealed a previously unrecognized role for IL13RA1 in myocardial homeostasis and cardiac function, making it an important target for cardiovascular research. To investigate cardiac pathophysiology using IL13RA1 Antibody, FITC conjugated, researchers can employ the following approaches:

• Expression Analysis in Normal vs. Pathological Samples:

  • Studies have shown that IL13RA1, together with the complementary type-2 IL-4 receptor chain IL4RA, is significantly downregulated in the hearts of patients with heart failure compared to normal hearts .

  • Researchers can use the FITC-conjugated antibody for flow cytometry or immunofluorescence microscopy to quantify and localize IL13RA1 expression in cardiac tissue samples from normal subjects versus various cardiovascular disease models.

• Functional Studies:

  • Research has demonstrated that IL13RA1 deficiency in mice leads to severe myocardial dysfunction and dyssynchrony (left ventricular ejection fraction 29.7±9.9 versus 45.0±8.0; P=0.004, left ventricular end-diastolic diameter 4.2±0.2 versus 3.92±0.3; P=0.03) .

  • The antibody can be used to correlate IL13RA1 expression levels with functional cardiac parameters in experimental models.

• Pathway Analysis:

  • Bioinformatic analysis of IL13RA1-deficient mouse hearts indicated that IL13RA1 regulates critical pathways beyond immune function, including extracellular matrix organization (normalized enrichment score=1.90; false discovery rate q=0.005) and glucose metabolism (normalized enrichment score=−2.36; false discovery rate q=0) .

  • Combining IL13RA1 staining with markers for these pathways can provide insights into the mechanistic roles of this receptor in cardiac pathophysiology.

• Fibrosis Assessment:

  • IL13RA1 deficiency has been associated with reduced collagen deposition under both normal and pressure-overload conditions .

  • Researchers can correlate IL13RA1 expression patterns with markers of fibrosis and extracellular matrix remodeling.

This approach provides a comprehensive framework for investigating the role of IL13RA1 in cardiac function and pathophysiology, potentially identifying new therapeutic targets for heart disease.

What approaches can be used to investigate the differential roles of IL-13 and IL-4 signaling through IL13RA1?

Investigating the differential roles of IL-13 and IL-4 signaling through IL13RA1 requires sophisticated experimental approaches that distinguish between type I and type II IL-4 receptor signaling pathways. Based on research findings, the following methodological approaches are recommended:

• Receptor Complex Characterization:

  • IL-4 can signal through both type I (IL4RA/IL2RG) and type II (IL4RA/IL13RA1) receptors, while IL-13 signals exclusively through type II receptors .

  • Use co-immunoprecipitation combined with immunofluorescence using the FITC-conjugated IL13RA1 antibody to visualize receptor complex formation under different stimulation conditions.

• Conditional Knockout Systems:

  • Studies using Il13ra1−/− mice have demonstrated that IL-13RA1 is essential for certain IL-13-induced responses but not for all IL-4-induced effects .

  • Research has shown that Th2 and IgE responses to T cell-dependent antigens are IL-13RA1-independent, while baseline IgE production requires IL-13RA1 .

  • Design experiments comparing wild-type, IL13RA1-deficient, and IL4RA-deficient models to parse out specific receptor contributions.

• Pathway-Specific Analysis:

  • Global expression profiling of lungs from mice stimulated with allergen or IL-4 has demonstrated that marker genes of alternatively activated macrophages are differentially regulated by the type I and type II IL-4R .

  • Use the FITC-conjugated antibody in conjunction with scRNA-seq or multi-parameter flow cytometry to identify cell populations responding to IL-13 stimulation.

• Functional Readouts:

  • Different downstream effects have been attributed to IL-13RA1 signaling, including:

    • Airway resistance and mucus production

    • TGF-β and eotaxin production

    • Regulation of alternatively activated macrophage gene expression

  • Design assays measuring these specific outcomes to differentiate between signaling pathways.

• Chemokine Response Analysis:

  • Research has demonstrated that IL-13 strongly induces chemokine expression (CCL2, CCL17, CCL11, and CCL24) in wild-type mice but not in Il13ra1−/− mice .

  • These differential responses can be used to distinguish IL-13-specific signaling from overlapping IL-4 effects.

Through these approaches, researchers can delineate the specific contributions of IL-13 and IL-4 signaling through the IL13RA1 receptor, providing insights into their distinct roles in health and disease.

What are common challenges with IL13RA1 Antibody, FITC conjugated and how can they be addressed?

When working with IL13RA1 Antibody, FITC conjugated, researchers may encounter several challenges that can impact experimental outcomes. Here are common issues and recommended solutions:

• Low Signal Intensity:

  • Challenge: FITC has relatively lower fluorescence intensity compared to some newer fluorophores and is susceptible to photobleaching.

  • Solution: Optimize antibody concentration through titration experiments (typically starting with 2-15 μg/mL) . Store samples protected from light and minimize exposure during imaging. Consider using anti-fading mounting media for immunofluorescence applications.

• High Background/Non-specific Binding:

  • Challenge: Polyclonal antibodies may exhibit higher background than monoclonals.

  • Solution: Implement more stringent blocking protocols using 5-10% normal serum from the same species as the secondary antibody. Include additional blocking agents such as BSA or casein. For flow cytometry, include appropriate Fc receptor blocking reagents and perform careful compensation for FITC autofluorescence .

• Poor Epitope Accessibility in Fixed Tissues:

  • Challenge: Formaldehyde fixation can mask epitopes.

  • Solution: Optimize antigen retrieval methods. For IL13RA1 detection in heart tissue, heat-induced epitope retrieval using a basic antigen retrieval reagent has been successful .

• Cross-Reactivity Concerns:

  • Challenge: Polyclonal antibodies might recognize related proteins.

  • Solution: Always include appropriate negative controls (isotype controls and/or tissues known to be negative for IL13RA1). Validate specificity using Il13ra1−/− tissues or cells if available .

• Storage and Stability Issues:

  • Challenge: FITC conjugates may lose activity with improper storage.

  • Solution: Store at -20°C or -80°C and avoid repeated freeze-thaw cycles . Consider aliquoting the antibody upon receipt.

• Variable Expression Levels:

  • Challenge: IL13RA1 expression may vary significantly between tissues and conditions.

  • Solution: Optimize exposure settings for each experimental condition. Research has shown variable expression in different tissues and downregulation in certain pathological conditions such as heart failure .

By implementing these optimization strategies, researchers can enhance the reliability and sensitivity of experiments using IL13RA1 Antibody, FITC conjugated.

How should researchers interpret IL13RA1 expression patterns in the context of cardiac disease?

Interpreting IL13RA1 expression patterns in cardiac disease requires careful consideration of multiple factors based on recent research findings:

• Expression Level Changes:

  • Studies have demonstrated that IL13RA1, together with the complementary type-2 IL-4 receptor chain IL4RA, is significantly downregulated in the hearts of patients with end-stage heart failure compared to normal donor hearts .

  • Researchers should quantify expression changes using appropriate controls and normalization strategies. Consider both protein level (using the FITC-conjugated antibody) and mRNA expression analyses for comprehensive assessment.

• Cellular Localization:

  • IL-13RA1 has been detected in the cytoplasm of myocardial cells using immunohistochemistry .

  • When interpreting immunofluorescence data, researchers should carefully document subcellular localization patterns and potential changes in distribution that may occur during disease progression.

• Correlation with Cardiac Function:

  • Research has established that Il13ra1-deficient mice develop severe myocardial dysfunction and dyssynchrony compared to wild-type mice, with significantly reduced left ventricular ejection fraction (29.7±9.9 versus 45.0±8.0; P=0.004) and increased left ventricular end-diastolic diameter (4.2±0.2 versus 3.92±0.3; P=0.03) .

  • When analyzing human samples, researchers should correlate IL13RA1 expression levels with functional cardiac parameters and clinical outcomes.

• Pathway Integration:

  • Bioinformatic analysis has revealed that IL13RA1 regulates critical pathways beyond the immune system, including:

    • Extracellular matrix organization (normalized enrichment score=1.90; false discovery rate q=0.005)

    • Glucose metabolism (normalized enrichment score=−2.36; false discovery rate q=0)

  • Researchers should interpret IL13RA1 expression changes in the context of these broader pathway alterations rather than in isolation.

• Relationship to Fibrosis:

  • IL13RA1 deficiency has been associated with reduced collagen deposition under both normal and pressure-overload conditions .

  • When analyzing cardiac samples, researchers should correlate IL13RA1 expression with markers of fibrosis and extracellular matrix remodeling to understand functional implications.

These considerations provide a framework for interpreting IL13RA1 expression data in cardiac disease, potentially identifying new therapeutic targets for heart disease treatment.

What are the key considerations when analyzing IL13RA1 expression in allergic and inflammatory conditions?

When analyzing IL13RA1 expression in allergic and inflammatory conditions, researchers should consider several important factors based on established research findings:

By considering these factors, researchers can develop a more nuanced interpretation of IL13RA1 expression data in allergic and inflammatory conditions, potentially leading to more targeted therapeutic approaches.

What are the emerging roles of IL13RA1 beyond traditional immune functions?

Recent research has uncovered several unexpected roles for IL13RA1 beyond its traditional functions in immune regulation, opening new avenues for investigation:

• Cardiac Homeostasis and Function:

  • Studies have revealed a previously unrecognized protective, regulatory role of IL13RA1 in myocardial homeostasis, metabolism, and repair .

  • Il13ra1-deficient mice develop severe myocardial dysfunction and dyssynchrony compared to wild-type mice, suggesting that IL13RA1 signaling is essential for normal cardiac function .

  • This finding represents a paradigm shift in understanding the role of this receptor beyond immune regulation.

• Extracellular Matrix Regulation:

  • Bioinformatic analysis of IL13RA1-deficient mouse hearts has demonstrated that IL13RA1 regulates extracellular matrix pathways (normalized enrichment score=1.90; false discovery rate q=0.005) .

  • IL13RA1 deficiency is associated with reduced collagen deposition under both normal and pressure-overload conditions, suggesting a direct role in tissue remodeling and fibrosis regulation .

• Metabolic Regulation:

  • IL13RA1 appears to regulate glucose metabolism pathways (normalized enrichment score=−2.36; false discovery rate q=0) in cardiac tissue .

  • This finding suggests potential implications for metabolic disorders and cardiac energetics.

• Differential Regulation of Inflammatory Pathways:

  • Research has identified an unexpected CCR3- and IL13RA1-independent pathway for lung eosinophilia, challenging previous assumptions about IL13RA1's role in allergic inflammation .

  • This discovery suggests more complex regulatory networks than previously appreciated.

• Autoregulatory Mechanisms:

  • Contrary to established paradigms, Il13ra1−/− mice exhibit elevated circulating soluble IL-13Rα2 and increased IL-13 levels, suggesting previously unrecognized feedback mechanisms .

These emerging roles highlight IL13RA1 as a multifunctional receptor involved in diverse physiological processes beyond traditional immune functions. Future research directions may include:

• Investigating IL13RA1 as a therapeutic target for heart failure and cardiomyopathies

• Exploring the role of IL13RA1 in metabolic disorders

• Examining tissue-specific functions of IL13RA1 in different organ systems

• Developing targeted therapies that modulate specific aspects of IL13RA1 signaling

These findings provide a foundation for expanded research into the diverse functions of this important receptor.

How might recent findings about IL13RA1 inform therapeutic approaches for inflammatory and cardiac diseases?

Recent discoveries about IL13RA1's functions have significant implications for developing new therapeutic strategies for both inflammatory and cardiac diseases:

These findings collectively suggest that modulating IL13RA1 signaling represents a promising therapeutic direction for multiple diseases, but successful approaches will require careful consideration of its diverse physiological roles and pathway-specific effects.

What controls are essential when using IL13RA1 Antibody, FITC conjugated in experimental settings?

Implementing appropriate controls is critical for ensuring the validity and interpretability of experiments using IL13RA1 Antibody, FITC conjugated. The following controls should be considered:

• Specificity Controls:

  • Isotype Control: Include a FITC-conjugated rabbit IgG isotype control at the same concentration as the primary antibody to assess non-specific binding .

  • Genetic Negative Control: When available, tissue or cells from Il13ra1−/− mice provide the ideal negative control for antibody specificity .

  • Blocking Peptide Control: Pre-incubation of the antibody with excess target peptide should abolish specific staining.

  • Primary Antibody Omission: For immunohistochemistry, samples processed without the primary antibody help identify background from secondary detection systems .

• Expression Controls:

  • Positive Tissue Control: Include tissues known to express IL13RA1, such as human granulocytes, skin, or heart tissue .

  • Negative Tissue Control: Include tissues with minimal IL13RA1 expression as determined by previous research.

  • Range of Expression Standards: When quantifying expression levels, include samples with known high, medium, and low expression levels of IL13RA1.

• Technical Controls:

  • Fluorescence Compensation Controls: For multi-parameter flow cytometry, single-stained controls are essential to correct for spectral overlap.

  • Autofluorescence Control: Unstained samples help establish baseline autofluorescence, particularly important for FITC which can overlap with natural tissue fluorescence.

  • Fixation Controls: When comparing fixed versus unfixed samples, include controls to assess the impact of fixation on epitope recognition.

• Experimental Design Controls:

  • Dose-Response Controls: For functional studies, include a range of antibody concentrations to establish optimal working concentrations.

  • Time-Course Controls: For dynamic processes, include multiple time points to capture temporal changes in IL13RA1 expression.

  • Treatment Controls: When assessing the effect of treatments on IL13RA1 expression, include appropriate vehicle controls.

• Validation Across Methods:

  • Orthogonal Validation: Confirm key findings using alternative methods (e.g., verify flow cytometry results with Western blotting or qPCR).

  • Antibody Validation: Consider using multiple antibodies targeting different epitopes of IL13RA1 to confirm expression patterns.

How can researchers design experiments to investigate IL13RA1's dual role in cardiac function and inflammatory responses?

Designing experiments to investigate IL13RA1's dual role in cardiac function and inflammatory responses requires careful consideration of multiple factors. The following experimental approach is recommended:

• Integrated Multi-System Models:

  • Challenge: Traditional models often focus on either cardiac function or inflammation in isolation.

  • Approach: Design experimental models that simultaneously assess cardiac parameters and inflammatory responses, such as:

    • Pressure overload models (e.g., transverse aortic constriction) in wild-type versus Il13ra1−/− mice, measuring both cardiac function and inflammatory markers

    • Allergen challenge models with cardiac function assessment in wild-type versus Il13ra1−/− mice

    • Combined models of cardiac injury and allergic challenge to assess potential interactions

• Cell Type-Specific Analysis:

  • Challenge: IL13RA1 may have different functions in different cell types.

  • Approach:

    • Use cell type-specific conditional knockout models (e.g., cardiomyocyte-specific, fibroblast-specific, or macrophage-specific Il13ra1 deletion)

    • Employ multi-parameter flow cytometry with FITC-conjugated IL13RA1 antibody to assess expression in different cell populations simultaneously

    • Perform single-cell RNA sequencing with targeted analysis of IL13RA1 and related pathway components

• Temporal Dynamics Assessment:

  • Challenge: IL13RA1's role may change during disease progression.

  • Approach:

    • Design longitudinal studies with multiple sampling timepoints

    • Implement inducible knockout systems to delete Il13ra1 at different disease stages

    • Use intravital imaging with fluorescently labeled antibodies to track receptor dynamics in real-time

• Pathway Dissection Experiments:

  • Challenge: IL13RA1 regulates multiple downstream pathways.

  • Approach:

    • Perform parallel assessment of multiple pathways:

      • Extracellular matrix organization (normalized enrichment score=1.90)

      • Glucose metabolism (normalized enrichment score=−2.36)

      • Inflammatory cytokine and chemokine production

    • Use pathway-specific inhibitors to determine which IL13RA1-dependent effects are mediated by which signaling cascades

• Translational Validation:

  • Challenge: Animal findings may not fully translate to human disease.

  • Approach:

    • Correlate findings from animal models with human samples

    • Analyze IL13RA1 expression in paired cardiac and immune cells from the same patients

    • Develop humanized mouse models expressing human IL13RA1 variants

• Intervention Testing:

  • Challenge: Understanding whether targeting IL13RA1 has therapeutic potential.

  • Approach:

    • Test IL13RA1 agonists in heart failure models

    • Evaluate IL13RA1 antagonists in allergic inflammation models

    • Assess the cardiac effects of established anti-IL-13 therapies used for asthma and atopic dermatitis

By implementing this comprehensive experimental approach, researchers can develop a more nuanced understanding of IL13RA1's dual role in cardiac function and inflammatory responses, potentially leading to novel therapeutic strategies that optimize effects across multiple physiological systems.