EPHA6 Antibody, FITC conjugated

What is EphA6 and why is it significant in cancer research?

EphA6 (Ephrin type-A receptor 6) is a receptor tyrosine kinase belonging to the Eph family that plays important roles in various biological functions including cancer development and progression. It has been identified as a potentially novel pro-metastatic gene in prostate cancer, with consistent overexpression observed in lymph node metastatic cell lines. Research has demonstrated that EphA6 contributes to cancer progression through multiple mechanisms including promotion of angiogenesis and enhancement of invasive properties. EphA6 expression is significantly higher in prostate cancer tumor tissues compared to benign tissues, and its expression positively correlates with vascular invasion, neural invasion, PSA levels, and TNM staging in prostate cancer cases . These characteristics make EphA6 a promising target for cancer metastasis therapy and an important focus for ongoing oncological research.

What applications is the EPHA6 Antibody, FITC conjugated suitable for?

EPHA6 Antibody, FITC conjugated has been validated for several research applications:

When using this antibody for any application, researchers should perform optimization studies to determine the ideal concentration for their specific experimental setup. The antibody has shown consistent results across these applications when used at the recommended dilutions, though laboratory-specific optimization may enhance performance .

What species reactivity does the EPHA6 Antibody, FITC conjugated demonstrate?

The reactivity profile of EPHA6 Antibody, FITC conjugated varies across manufacturers but typically includes:

When working with species not listed or confirmed, researchers should perform preliminary validation experiments to confirm cross-reactivity. The broad species reactivity of some products makes this antibody versatile for comparative studies across different animal models .

What are the optimal storage and handling protocols for EPHA6 Antibody, FITC conjugated?

For optimal performance and longevity of EPHA6 Antibody, FITC conjugated, researchers should follow these storage and handling guidelines:

• Long-term storage: Store at -20°C or -80°C in the dark to preserve FITC fluorescence .

• Reconstitution stability: When reconstituted in sterile pH 7.4 0.01M PBS or recommended diluent, the antibody remains stable for at least two weeks at 2-4°C .

• Avoid repeated freeze/thaw cycles: These can deteriorate antibody quality and reduce binding efficacy .

• Working solution preparation: Prepare fresh dilutions on the day of experiment when possible.

• Light sensitivity: FITC is photosensitive - minimize exposure to light during handling, storage, and experimentation to prevent photobleaching.

• Buffer compatibility: Most products are supplied in a buffer containing glycerol (typically 50%) and PBS with preservatives like Proclin 300 (0.03%) .

Following these guidelines ensures maximum antibody performance and extends the usable lifespan of the reagent across multiple experiments.

How can EPHA6 Antibody, FITC conjugated be used to study cancer progression and metastasis?

EPHA6 Antibody, FITC conjugated provides researchers with powerful tools to investigate EphA6's role in cancer progression through several advanced applications:

• Metastatic cell identification: The antibody can be used to visualize and quantify EphA6-expressing cells in metastatic sites via immunofluorescence microscopy. Research has demonstrated that EphA6 knockdown significantly decreases the incidence of metastases to local draining lymph nodes and lungs in prostate cancer xenograft models .

• Co-localization studies: Using multicolor immunofluorescence approaches, researchers can investigate EphA6 co-localization with other metastasis-associated proteins to elucidate interaction networks.

• Dynamic expression analysis: Studies have shown that EphA6 expression increases in metastatic derivative cancer cells compared to their parental lines. The FITC-conjugated antibody enables visualization of this differential expression in tissue samples and cell cultures .

• Mechanism investigation: Research has identified that EphA6 contributes to metastasis through multiple mechanisms including enhanced matrix degradation and invasion. Using the antibody in combination with functional assays can help elucidate these mechanisms .

• Clinical correlation studies: Immunofluorescence analysis of patient samples has revealed that EphA6 protein is strongly expressed in primary prostate cancer tumor tissues while minimal in adjacent non-tumor tissues, with significant positive correlation to vascular invasion, neural invasion, PSA level, and TNM staging .

These applications highlight how the FITC-conjugated antibody serves as an essential tool in investigating the molecular mechanisms of cancer metastasis mediated by EphA6.

What is the role of EphA6 in angiogenesis and how can the antibody help investigate this process?

EphA6 has been identified as a positive regulator of angiogenesis, and EPHA6 Antibody, FITC conjugated provides valuable insights into this process:

• Mechanism of action: Research demonstrates that EphA6 knockdown decreases tube formation in vitro and reduces angiogenesis in vivo. The FITC-conjugated antibody can be used to visualize EphA6 expression in endothelial cells and tumor vasculature, helping elucidate its role in neovascularization .

• Vascular density quantification: Studies show that EphA6 knockdown decreases microvascular density (MVD) in tumor tissues. Researchers can use the antibody to quantify MVD through immunofluorescence analysis of tissue sections .

• Paracrine signaling investigation: Conditioned media from EphA6 knockdown cancer cells shows decreased ability to induce endothelial cell tube formation, suggesting paracrine regulation of angiogenesis. The antibody can help track EphA6-expressing cancer cells and their interaction with the vascular microenvironment .

• Co-staining with vascular markers: For comprehensive analysis, researchers can perform co-immunofluorescence with the FITC-conjugated EPHA6 antibody and endothelial markers (CD31, CD34) to investigate the relationship between EphA6 expression and vessel formation in tumors.

• Mechanistic pathway analysis: Evidence suggests EphA6 may promote angiogenesis through interaction with the PI3K/AKT pathway. The antibody can be used alongside phospho-specific antibodies to elucidate signaling mechanisms .

These applications demonstrate how EPHA6 Antibody, FITC conjugated serves as a critical tool in dissecting the complex role of EphA6 in pathological angiogenesis.

How does EphA6 interact with the PI3K/AKT signaling pathway in cancer progression?

Research has revealed important connections between EphA6 and the PI3K/AKT pathway that can be further investigated using EPHA6 Antibody, FITC conjugated:

• Downstream effector regulation: Genome-wide gene expression analysis following EphA6 knockdown identified several genes regulated by EphA6 that are either components of the PI3K/AKT pathway or associated with this pathway. The antibody can help visualize changes in EphA6 expression concurrent with alterations in pathway components .

• AKT modulation: Studies show that knockdown of EphA6 decreased AKT expression, a major component of the PI3K/AKT pathway. Researchers can use the antibody in conjunction with phospho-AKT antibodies to track this relationship .

• EIF5A2 connection: EIF5A2, an AKT target gene that promotes melanoma cell invasion, has decreased expression following EphA6 knockdown. Co-immunofluorescence studies with both markers can elucidate this relationship .

• PIK3IP1 regulation: The expression of PIK3IP1, a negative regulator of PI3K and suppressor of tumor development, increases after EphA6 knockdown. This suggests EphA6 may normally suppress this tumor suppressor .

• Spatial organization analysis: Using the FITC-conjugated antibody, researchers can visualize the subcellular localization of EphA6 relative to PI3K/AKT signaling components through high-resolution confocal microscopy.

The research findings suggest that EphA6 may exert its pro-metastatic effects at least partially through modulation of the PI3K/AKT signaling pathway, a relationship that can be further elucidated using fluorescently labeled antibodies in various imaging-based applications.

What controls should be included when using EPHA6 Antibody, FITC conjugated in immunofluorescence studies?

When designing immunofluorescence experiments with EPHA6 Antibody, FITC conjugated, researchers should incorporate the following controls:

• Positive control: Include known EphA6-expressing cells or tissues such as prostate cancer cell lines PC-3M or CWR22rv1, which have been documented to express EphA6 .

• Negative control: Include normal prostate epithelial cell lines like p69 or RWPE1, which have been shown to have undetectable EphA6 expression by Western blot analysis .

• Isotype control: Include a FITC-conjugated rabbit IgG (matching the host species and isotype of the EPHA6 antibody) at the same concentration to assess non-specific binding .

• Secondary antibody-only control: For experimental designs using secondary detection systems, include samples with secondary antibody only to identify potential background.

• Blocking peptide control: If available, pre-incubate the antibody with its specific immunogen peptide to confirm signal specificity.

• Autofluorescence control: Include unstained samples to assess natural tissue autofluorescence, particularly important when working with tissues containing lipofuscin or elastin.

• Signal validation using alternative detection: Compare FITC signal patterns with results from non-fluorescent detection methods (such as chromogenic IHC) to confirm specificity.

• Knockdown/knockout validation: When possible, include EphA6 knockdown samples (using verified shRNA constructs) as the gold standard for specificity validation .

Incorporating these controls ensures reliable data interpretation and helps distinguish true EphA6-specific signals from technical artifacts.

How can researchers optimize immunofluorescence protocols with EPHA6 Antibody, FITC conjugated?

Achieving optimal results with EPHA6 Antibody, FITC conjugated requires careful protocol optimization:

• Fixation optimization:

  • For cell lines: 4% paraformaldehyde for 15 minutes at room temperature preserves EPHA6 epitopes while maintaining cellular morphology

  • For tissues: 10% neutral buffered formalin with controlled fixation time (24-48 hours) followed by proper paraffin embedding

• Antigen retrieval methods:

  • Heat-induced epitope retrieval using citrate buffer (pH 6.0) for 20 minutes typically yields good results

  • For difficult samples, try Tris-EDTA buffer (pH 9.0) as an alternative

• Blocking optimization:

  • Use 5-10% normal serum (from species unrelated to primary antibody host) with 0.1-0.3% Triton X-100 for permeabilization

  • Include 1% BSA to reduce non-specific binding

• Antibody dilution optimization:

  • Start with manufacturer's recommended dilution range (1:50-200 for IF)

  • Perform a dilution series to identify optimal signal-to-noise ratio for specific samples

• Incubation conditions:

  • Extended incubation (overnight at 4°C) often yields better results than short incubations

  • Maintain consistent temperature and humidity throughout incubation

• Washing optimization:

  • Multiple (3-5) washes with PBS-T (PBS + 0.05% Tween-20)

  • Extend wash duration (5-10 minutes per wash) to reduce background

• Counterstain selection:

  • Choose nuclear counterstains that don't overlap with FITC spectrum (DAPI or Hoechst recommended)

  • For co-localization studies, select fluorophores with minimal spectral overlap with FITC (Alexa 555/594/647)

• Mounting media consideration:

  • Use anti-fade mounting media specifically formulated for fluorescence

  • Consider media with DAPI if nuclear counterstaining is needed

• Photobleaching prevention:

  • Minimize exposure to light during all steps

  • Store slides in the dark and image promptly after preparation

Following these optimization steps will help researchers achieve consistent, high-quality results when using EPHA6 Antibody, FITC conjugated for immunofluorescence applications.

What sample preparation techniques yield optimal results for detecting EphA6 in tissue samples?

Effective detection of EphA6 in tissue samples requires careful attention to sample preparation:

• Tissue collection and fixation:

  • Collect tissues rapidly after excision to minimize protein degradation

  • Fix tissues in 10% neutral buffered formalin for 24-48 hours (excessive fixation can mask epitopes)

  • Consider using PAXgene or other molecular fixatives for dual immunofluorescence/molecular analysis

• Processing and embedding:

  • Use standardized processing protocols with controlled temperature and reagent quality

  • Optimal embedding in high-quality paraffin prevents sectioning artifacts

  • Flash-frozen tissues may preserve antigenicity but require different sectioning approaches

• Sectioning parameters:

  • 4-5 μm sections provide optimal resolution for EphA6 detection

  • Mount sections on positively charged slides to prevent tissue loss during processing

  • Allow sections to air-dry thoroughly before storage or processing

• Pre-treatment protocols:

  • Deparaffinization must be complete (multiple xylene changes)

  • Hydration through graded alcohols should be thorough

  • Antigen retrieval optimization is critical - studies have shown heat-induced epitope retrieval with citrate buffer (pH 6.0) works well for EphA6 detection

• Permeabilization considerations:

  • For FFPE tissues: 0.2% Triton X-100 for 10 minutes typically provides adequate permeabilization

  • For frozen sections: 0.1% Triton X-100 is usually sufficient

  • Over-permeabilization can disrupt tissue morphology and increase background

• Autofluorescence reduction:

  • Treat sections with 0.1% Sudan Black B in 70% ethanol for 20 minutes to reduce autofluorescence

  • Commercial autofluorescence quenching reagents may also be effective

  • Photobleaching treatment with light exposure prior to antibody incubation can reduce background

• Blocking optimization:

  • Include both protein blocking (5-10% normal serum) and avidin-biotin blocking steps

  • Extend blocking time (1-2 hours) for tissues with high background potential

• Control tissue inclusion:

  • Process known positive tissues (prostate cancer samples) alongside test tissues

  • Include normal prostate epithelial tissue as negative/low expression control

These optimized sample preparation techniques help ensure reliable and reproducible detection of EphA6 in tissue specimens, enhancing the scientific validity of research findings.

How can researchers distinguish between specific and non-specific signals when using EPHA6 Antibody, FITC conjugated?

Differentiating genuine EphA6 signal from non-specific background is critical for accurate data interpretation:

• Pattern recognition:

  • Genuine EphA6 staining typically shows membrane localization with some cytoplasmic signal

  • Non-specific staining often appears as diffuse cytoplasmic signal or nuclear staining

  • Compare observed patterns with published EphA6 localization studies

• Signal validation approaches:

  • Perform parallel staining with a second EphA6 antibody targeting a different epitope

  • Compare FITC signal with mRNA expression data from the same samples

  • Validate using functional approaches (e.g., shRNA knockdown decreases signal)

• Control comparisons:

  • Compare signal intensity between positive controls (prostate cancer cells) and negative controls (normal prostate epithelial cells)

  • Isotype control should show minimal background at the same exposure settings

• Technical considerations:

  • Autofluorescence can be distinguished by its presence in multiple channels

  • Edge artifacts typically appear at tissue borders and have distinct morphology

  • Non-specific binding often occurs in necrotic areas or regions with high protein content

• Quantitative assessment:

  • Establish signal-to-noise ratio thresholds based on control samples

  • Consider automated analysis tools that can distinguish specific signal based on intensity and localization parameters

  • Compare signal distribution across different cell types within the same sample (internal control)

• Signal characteristics matrix:

Using these approaches systematically helps researchers confidently identify genuine EphA6 expression and distinguish it from technical artifacts.

What are potential causes of inconsistent results when using EPHA6 Antibody, FITC conjugated?

When researchers encounter variability in results using EPHA6 Antibody, FITC conjugated, several factors may be responsible:

• Antibody-related issues:

  • FITC photobleaching from improper storage or excessive light exposure

  • Antibody degradation from repeated freeze-thaw cycles

  • Lot-to-lot variability in antibody production

  • Inadequate antibody concentration for specific application

• Sample preparation variables:

  • Inconsistent fixation procedures altering epitope availability

  • Variable antigen retrieval efficiency between experiments

  • Differences in tissue processing affecting tissue morphology

  • Irregular section thickness causing variability in signal intensity

• Technical procedure inconsistencies:

  • Variations in incubation times or temperatures

  • Inadequate washing leading to high background

  • Inconsistent blocking efficiency between experiments

  • Variable permeabilization affecting antibody penetration

• Biological variability:

  • Natural heterogeneity in EphA6 expression across different regions of the same tumor

  • Cell cycle-dependent fluctuations in receptor expression

  • Microenvironmental factors affecting EphA6 regulation

  • Treatment-induced changes in receptor expression or localization

• Instrument and imaging variables:

  • Inconsistent microscope settings between sessions

  • Variable exposure times affecting signal intensity

  • Differences in filter sets between microscopes

  • Software-based image processing variations

• Troubleshooting approach matrix:

Addressing these variables systematically can significantly improve reproducibility when working with EPHA6 Antibody, FITC conjugated in research applications.

How can researchers quantify EphA6 expression using FITC-conjugated antibodies?

Accurate quantification of EphA6 expression using FITC-conjugated antibodies requires robust methodological approaches:

• Image-based quantification methods:

  • Mean fluorescence intensity (MFI) measurement in defined regions of interest

  • Pixel intensity histograms to assess signal distribution

  • Membrane:cytoplasm signal ratio analysis for localization assessment

  • Threshold-based binary masks to identify positive versus negative cells

• Flow cytometry-based quantification:

  • Single-cell suspension preparation from tissues or cultures

  • Gating strategies to identify EphA6-positive populations

  • Comparative analysis with calibration beads for absolute quantification

  • Mean or median fluorescence intensity measurements for relative expression

• Standardization approaches:

  • Include calibration controls in each experiment (cells with known EphA6 expression levels)

  • Use internal controls (cells or regions with consistent expression) within samples

  • Apply background subtraction based on isotype controls

  • Normalize to reference markers or housekeeping proteins

• Advanced quantification techniques:

  • Co-localization coefficient calculation for interaction studies

  • Machine learning-based segmentation for complex tissue architecture

  • 3D reconstruction for volumetric assessment of expression patterns

  • Time-lapse imaging for dynamic expression changes

• Quantification workflow example:

  a. Capture multiple representative fields (minimum 5-10) using identical acquisition settings
  b. Apply flat-field correction to account for illumination heterogeneity
  c. Perform background subtraction based on negative control samples
  d. Define regions of interest (cells, tissues, or subcellular compartments)
  e. Measure integrated density or mean fluorescence intensity
  f. Apply statistical analysis to compare across experimental conditions

• Validation approaches:

  • Correlate fluorescence quantification with Western blot or qPCR data

  • Compare results from different quantification methods for consistency

  • Assess technical reproducibility through repeated measurements

  • Perform biological replicates to account for sample variability

These quantification approaches enable researchers to obtain reliable, reproducible measurements of EphA6 expression patterns, facilitating meaningful comparisons across experimental conditions and accurate interpretation of biological significance.

How does EphA6 expression correlate with clinical parameters in cancer patients?

EphA6 expression demonstrates significant correlations with several clinical parameters, providing important diagnostic and prognostic insights:

• Expression patterns in normal versus malignant tissue:

  • Studies using immunohistochemistry have demonstrated that EphA6 protein is strongly expressed in primary prostate cancer tissues while minimal expression is detected in adjacent non-tumor tissues .

  • The number of cells positive for EphA6 is significantly higher in primary cancer tissues than in matched adjacent non-tumor tissues .

• Correlation with clinical characteristics:

  • Research has identified positive correlations between EphA6 expression and several clinical parameters:

    • Vascular invasion status

    • Neural invasion status

    • PSA levels

    • TNM staging

• Relationship with Gleason score:

  • Interestingly, studies have not shown significant correlation between EphA6 expression in tumors and Gleason scores, potentially because samples analyzed typically started from Gleason score 6, representing already aggressive prostate cancer .

• Metastatic potential relationship:

  • EphA6 expression is consistently higher in lymph node metastatic cell lines compared to their parental lines, suggesting association with metastatic potential .

  • Samples from EphA6 knockdown experimental groups showed significantly decreased incidence of metastases to local draining lymph nodes and lungs compared to control groups in animal models .

• Clinical significance table:

These clinical correlations suggest that EphA6 may serve as a potential biomarker for cancer aggressiveness and metastatic potential, with possible applications in patient stratification and therapeutic decision-making.

What are the potential therapeutic implications of targeting EphA6 in cancer?

Research into EphA6 biology has revealed several promising therapeutic implications:

• Anti-metastatic potential:

  • Experimental knockdown of EphA6 using shRNA significantly reduces prostate cancer metastasis to lymph nodes and lungs in animal models, suggesting that therapeutic targeting of EphA6 could limit metastatic spread .

  • The strong association between EphA6 expression and metastatic potential indicates that inhibiting EphA6 function may represent a viable strategy for preventing or reducing metastasis in high-risk patients .

• Anti-angiogenic effects:

  • EphA6 knockdown decreases tube formation in vitro and reduces angiogenesis in vivo, suggesting that EphA6-targeted therapies might function partly through inhibition of tumor vascularization .

  • Tumors with EphA6 knockdown present reduced microvascular density, indicating that EphA6 inhibition could complement existing anti-angiogenic therapies .

• Pathway modulation opportunities:

  • The connection between EphA6 and the PI3K/AKT pathway suggests that targeting EphA6 could modulate this critical oncogenic signaling pathway .

  • EphA6 regulates multiple genes involved in metastasis, including PIK3IP1, AKT1, and EIF5A2, providing multiple downstream mechanisms for therapeutic benefit .

• Combinatorial therapy potential:

  • The role of EphA6 in both invasion and angiogenesis suggests that targeting this receptor could enhance the efficacy of existing treatments by addressing multiple aspects of tumor biology simultaneously.

  • Targeting EphA6 in conjunction with conventional therapies might provide synergistic benefits, particularly for aggressive, metastatic cancers.

• Therapeutic development considerations:

  • Antibody-based therapies (including antibody-drug conjugates) targeting EphA6 represent a potential therapeutic approach.

  • Small molecule inhibitors targeting EphA6 kinase activity could block downstream signaling.

  • Gene therapy approaches to downregulate EphA6 expression might recapitulate the beneficial effects seen in experimental knockdown studies.

• Biomarker applications:

  • EphA6 expression analysis using FITC-conjugated antibodies could help identify patients most likely to benefit from EphA6-targeted therapies.

  • Monitoring changes in EphA6 expression might serve as a pharmacodynamic marker for treatment efficacy.

These findings collectively support the potential therapeutic value of targeting EphA6 in cancer, particularly for preventing or treating metastatic disease, with FITC-conjugated antibodies serving important roles in both research and potential clinical applications.

What emerging technologies might enhance EphA6 detection and functional analysis?

Several cutting-edge technologies show promise for advancing EphA6 research beyond current methodologies:

• Advanced imaging approaches:

  • Super-resolution microscopy techniques (STED, PALM, STORM) can provide nanoscale visualization of EphA6 clustering and receptor interactions using FITC-conjugated antibodies

  • Light sheet microscopy enables 3D visualization of EphA6 distribution in intact tissues with reduced photobleaching

  • Intravital microscopy allows real-time tracking of EphA6-expressing cells in living organisms

• Single-cell analysis technologies:

  • Single-cell RNA sequencing combined with protein analysis can correlate EphA6 expression with global transcriptional programs

  • Mass cytometry (CyTOF) enables simultaneous detection of EphA6 with dozens of other markers without fluorescence overlap concerns

  • Digital spatial profiling allows quantitative assessment of EphA6 expression with spatial context in tissue microenvironments

• Functional genomics approaches:

  • CRISPR-Cas9 screening can identify synthetic lethal interactions with EphA6 in cancer cells

  • CRISPR activation/repression systems allow precise modulation of EphA6 expression levels

  • Base editing techniques enable introduction of specific EphA6 mutations to study functional consequences

• Protein interaction analysis:

  • Proximity labeling methods (BioID, APEX) can identify novel EphA6 interacting proteins in living cells

  • FRET-based approaches using FITC-conjugated antibodies can detect protein-protein interactions involving EphA6

  • Protein complementation assays provide functional readouts of receptor activation states

• Translational research tools:

  • Patient-derived organoids enable testing of EphA6-targeting approaches in models that better recapitulate human disease

  • Humanized mouse models allow investigation of EphA6 biology in more clinically relevant contexts

  • Liquid biopsy approaches may enable non-invasive monitoring of EphA6-expressing circulating tumor cells

• Artificial intelligence applications:

  • Machine learning algorithms can improve image analysis and quantification of EphA6 staining patterns

  • Deep learning approaches may identify subtle correlations between EphA6 expression and clinical outcomes

  • AI-driven drug discovery could accelerate development of EphA6-targeting therapeutics

These emerging technologies promise to deepen our understanding of EphA6 biology and accelerate translation of basic research findings into clinical applications, with FITC-conjugated antibodies continuing to play important roles in many of these advanced approaches.

What unresolved questions remain regarding EphA6 biology and function?

Despite significant advances in EphA6 research, several important questions remain unanswered:

• Signaling mechanism specificity:

  • How does EphA6 signaling differ from other Eph family members?

  • What determines whether EphA6 promotes forward or reverse signaling in specific contexts?

  • Which ephrin ligands preferentially interact with EphA6 in different tissue types?

• Regulation of expression:

  • What transcriptional, epigenetic, and post-transcriptional mechanisms control EphA6 expression?

  • How is EphA6 expression dynamically regulated during cancer progression and metastasis?

  • What factors drive the overexpression of EphA6 observed in prostate cancer tissues?

• Pathway interactions:

  • Beyond the identified connections to PI3K/AKT, what other signaling pathways interact with EphA6?

  • How does EphA6 specifically regulate the identified downstream genes (PIK3IP1, AKT1, EIF5A2)?

  • What is the precise molecular mechanism by which EphA6 contributes to angiogenesis?

• Functional heterogeneity:

  • Does EphA6 function differently across cancer types or even within different regions of the same tumor?

  • How does the tumor microenvironment influence EphA6 activity and function?

  • Are there cancer stem cell-specific roles for EphA6 in maintaining tumor-initiating populations?

• Therapeutic targeting challenges:

  • What approaches most effectively inhibit EphA6 function in vivo?

  • How can potential off-target effects of EphA6 inhibition be mitigated?

  • Will resistance mechanisms emerge following EphA6-targeted therapy?

• Clinical relevance questions:

  • Beyond prostate cancer, which other tumor types show EphA6 dependence?

  • Can EphA6 expression serve as a biomarker for patient stratification in clinical trials?

  • Does EphA6 expression predict response to specific therapeutic regimens?

• Methodological limitations:

  • How can we better distinguish EphA6-specific effects from those shared with other Eph family members?

  • What are the optimal models for studying EphA6 function in complex tissue environments?

  • How can we improve the specificity and sensitivity of tools for detecting EphA6 activation states?

Addressing these unresolved questions will require multidisciplinary approaches and innovative experimental designs, with FITC-conjugated antibodies remaining valuable tools for visualizing and tracking EphA6 expression across diverse experimental systems.