IRC11 Antibody

What is Integrin Alpha 11 and what biological systems express it?

Integrin Alpha 11 is a transmembrane receptor that forms heterodimers with beta subunits, particularly beta-1. It functions primarily in cell-collagen interactions and is implicated in tissue fibrosis pathways. The protein is expressed in various tissues, with notable detection in human heart tissue and hepatic stellate cells (HSCs) as demonstrated by Western blot analyses. During fibrotic processes, Integrin Alpha 11 expression increases significantly in activated HSCs, making it an important marker and potential therapeutic target in fibrosis research .

What detection methods are validated for Integrin Alpha 11 antibody?

The Integrin Alpha 11 antibody has been validated for multiple applications with specific methodological considerations:

• Western Blot: Effective under non-reducing conditions using Immunoblot Buffer Group 1, detecting bands at approximately 150 kDa

• Direct ELISA: Successfully detects human and mouse Integrin alpha 11

• Immunofluorescence: Enables visualization of protein localization in cells and tissues

• Flow Cytometry: Validated for detection of surface expression

For Western blot applications specifically, probing with 2 μg/mL of antibody followed by HRP-conjugated Anti-Rat IgG Secondary Antibody produces optimal results .

How can I confirm antibody specificity for Integrin Alpha 11?

Specificity confirmation requires multiple complementary approaches:

• Cross-reactivity testing: The antibody has been shown not to cross-react with recombinant human Integrin alpha 2 in direct ELISAs

• Molecular weight verification: Confirm detection at the expected molecular weight (~150 kDa)

• Positive controls: Use tissues with known expression like human heart tissue or HeLa cells

• Knockdown validation: siRNA experiments targeting Integrin Alpha 11 should show corresponding reduction in antibody signal

• Comparing multiple antibody clones: Using antibodies recognizing different epitopes to confirm consistent detection patterns

How does Integrin Alpha 11 contribute to liver fibrosis mechanisms?

Integrin Alpha 11 plays a crucial role in liver fibrosis through several mechanisms:

• Regulation of pro-fibrotic signaling proteins: Research demonstrates that knockdown of Integrin Alpha 11 (Itga11) leads to diminished expression of MYL9 and YAP-1, key mediators of fibrosis

• Cytoskeletal reorganization: Loss of Integrin beta-1 (binding partner of alpha 11) causes HSCs to adopt a rounded, inactivated appearance

• Transcriptional regulation: Itga11 knockdown using two different siRNA oligos shows corresponding reduction in Myl9 transcripts, indicating a regulatory role in gene expression

• YAP-1 phosphorylation modulation: Integrin signaling affects the phosphorylation status of YAP-1, with disruption leading to increased inactive phosphorylated YAP (PYAP)

The following data table summarizes experimental findings from Integrin Alpha 11 knockdown studies:

What methodological approaches are effective for studying Integrin Alpha 11 in fibrosis models?

When investigating Integrin Alpha 11 in fibrosis research, several methodological approaches have proven effective:

• siRNA knockdown: Using multiple oligos targeting different regions of Itga11 mRNA to ensure specificity, followed by assessment of downstream effectors like MYL9 and YAP-1

• Genetic deletion models: Studying the effects of Integrin beta-1 deletion (Itgb1-null) provides insights into the function of the alpha-beta heterodimer

• Immunofluorescence localization: Determining subcellular localization of Integrin Alpha 11 and associated proteins like YAP-1

• Protein-protein interaction studies: Investigating the relationship between Integrin Alpha 11 and other fibrosis mediators

• Transcriptional analysis: Measuring mRNA levels of Itga11 and related genes following experimental manipulations

How does Integrin Alpha 11 function differ between normal and pathological conditions?

In normal tissues, Integrin Alpha 11 mediates cell-collagen interactions with controlled signaling. Under pathological conditions, particularly fibrosis, several important differences emerge:

• Expression levels: Significant upregulation in activated HSCs during fibrotic processes

• Signaling pathway activation: Enhanced activation of YAP-1 and MYL9 pathways

• ECM production promotion: Contributes to excessive collagen production through downstream signaling

• Cell morphology regulation: Helps maintain the activated phenotype of fibroblasts and HSCs

• Mechanosensing alterations: Affects cellular response to matrix stiffness, creating a feedback loop in fibrosis progression

What are optimal sample preparation conditions for Integrin Alpha 11 detection?

Successful detection requires careful sample preparation:

• Non-reducing conditions: Western blot detection is optimal under non-reducing conditions

• Buffer selection: Use of Immunoblot Buffer Group 1 is recommended for Western blot applications

• Cell lysis optimization: For cellular samples, complete lysis while preserving protein integrity is critical

• Fresh tissue handling: When using tissues like heart, rapid processing minimizes protein degradation

• Detection antibody concentration: 2 μg/mL has been validated for optimal signal-to-noise ratio in Western blots

How can I optimize antibody dilutions for different applications?

Antibody dilution optimization requires systematic testing:

• Application-specific ranges:

  • Western blot: Start with 1-2 μg/mL

  • ELISA: Initial testing at 0.5-1 μg/mL

  • Immunofluorescence: Begin with 5-10 μg/mL and adjust

• Titration approach: Test 3-4 dilutions spanning a 5-10 fold range

• Signal-to-noise assessment: Evaluate specific signal versus background at each dilution

• Sample-specific adjustments: Different sample types may require modified concentrations

• Incubation conditions: Consider adjusting time and temperature alongside concentration

How do I troubleshoot inconsistent detection of Integrin Alpha 11?

When facing detection challenges, consider these methodological approaches:

• Sample quality assessment: Verify protein integrity with a housekeeping protein control

• Blocking optimization: Test alternative blocking agents if background is high

• Epitope accessibility: If signal is weak, mild denaturation techniques may expose masked epitopes

• Antibody validation: Confirm antibody activity with a positive control sample

• Detection system sensitivity: For low abundance, consider switching to more sensitive detection systems like chemiluminescence with signal enhancement or amplification steps

How can ultra-sensitive detection methods be applied to Integrin Alpha 11 research?

While not specifically developed for Integrin Alpha 11, recent advances in ultra-sensitive detection technologies offer opportunities for enhanced research:

• Ultra-sensitive platforms comparison:

  • Meso Scale Discovery (MSD): Provides enhanced sensitivity over standard ELISA

  • Simoa HD-1: Offers single-molecule detection capability

  • Simoa Planar Array (SP-X): Achieves detection at sub-picogram levels

• Epitope binning importance: When developing high-sensitivity assays, screening antibodies from distinct epitope communities is critical for creating optimal capture-detection pairs

• Minimum required dilution (MRD): Determining optimal sample dilution through spike recovery and dilution linearity experiments

• Cross-species detection optimization: When studying models across species, validation of cross-reactivity and species-specific sensitivity is essential

What computational approaches might benefit Integrin Alpha 11 antibody research?

Recent developments in computational antibody design suggest promising applications:

• Deep learning models: Machine learning approaches can generate antibody sequences with favorable developability attributes

• Medicine-likeness screening: Computational assessment of antibody properties that resemble marketed antibody therapeutics

• In-silico antibody libraries: Generation of diverse antibody sequences with high humanness and developability

• Structural modeling: Prediction of antibody-antigen interactions to guide experimental design

• Epitope mapping: Computational approaches to identify optimal binding regions for antibody development

What experimental considerations are important when studying Integrin Alpha 11 in multi-protein complexes?

Studying Integrin Alpha 11 within its functional protein complexes requires specialized approaches:

• Co-immunoprecipitation optimization: Using appropriate detergents to maintain complex integrity while enabling antibody access

• Proximity ligation assays: For detecting in-situ protein-protein interactions involving Integrin Alpha 11

• Multiple epitope targeting: Using antibodies against different components of the complex simultaneously

• Native gel electrophoresis: Preserving protein-protein interactions during separation

• Functional assays: Correlating complex formation with downstream signaling events like YAP-1 nuclear translocation

How might Integrin Alpha 11 antibodies contribute to fibrosis therapeutic development?

The research suggests several promising therapeutic avenues:

• Target validation approaches: Using Integrin Alpha 11 antibodies to validate the protein as a therapeutic target through functional blocking studies

• Biomarker development: Employing antibodies to measure Integrin Alpha 11 levels as a biomarker of fibrotic disease progression

• Mechanism elucidation: Further defining the signaling pathway from Integrin Alpha 11 to YAP-1 and MYL9 to identify additional intervention points

• Humanized antibody development: Transitioning from research antibodies to potential therapeutic candidates

• Combination therapy assessment: Evaluating Integrin Alpha 11 targeting in combination with other anti-fibrotic approaches

What novel detection methods might enhance Integrin Alpha 11 research?

Emerging technologies that could advance Integrin Alpha 11 research include:

• Multiplex imaging: Simultaneous detection of Integrin Alpha 11 with multiple signaling partners

• Single-cell analysis: Examining heterogeneity of Integrin Alpha 11 expression within tissue populations

• In vivo imaging: Development of labeled antibodies for tracking Integrin Alpha 11 in animal models

• Mass cytometry: Adding Integrin Alpha 11 to CyTOF panels for comprehensive cellular profiling

• Spatial transcriptomics integration: Correlating protein detection with spatial gene expression patterns