CELSR1 Antibody

What is CELSR1 and what are its key structural features?

CELSR1 is an atypical cadherin protein with a calculated molecular weight of approximately 329.5 kDa . Structurally, CELSR1 contains multiple functional domains arranged in modules. The protein features cadherin domains (CADH1-8), which are essential for its adhesive functions, as well as a compact module consisting of CADH9 through GAIN domains that plays distinct roles in cellular processes . CELSR1 is also known by several alternative names including ADGRC1, CDHF9, FMI2, HFMI2, adhesion G protein-coupled receptor C1, and cadherin family member 9 . As a flamingo homolog (from Drosophila), CELSR1 serves as an important component in planar cell polarity pathways, particularly in developmental contexts .

How should researchers select an appropriate CELSR1 antibody for their experiments?

When selecting a CELSR1 antibody, researchers should consider several critical factors. First, determine the specific epitope recognition needed for your application - some antibodies target the extracellular domain while others recognize the cytoplasmic domain . For example, studies examining Celsr1 distribution in mouse sensory epithelia have successfully used antibodies against amino acids 2885-2985 in the cytoplasmic domain .

Second, evaluate reactivity with your species of interest. Available antibodies show reactivity with human, mouse, and rat CELSR1, though species cross-reactivity varies by product . Third, confirm application compatibility - different antibodies are validated for specific applications including ELISA, immunofluorescence (IF), western blot (WB), immunoprecipitation (IP), and immunohistochemistry (IHC) . Finally, consider antibody format (polyclonal versus monoclonal) based on your experimental needs; polyclonal antibodies may provide greater epitope coverage while monoclonals offer higher specificity .

What are the typical applications for CELSR1 antibodies in research settings?

CELSR1 antibodies are utilized across multiple experimental applications in research settings:

• Immunofluorescence/Immunocytochemistry: Visualizing the subcellular distribution of CELSR1, particularly at cell-cell junctions. This technique has been instrumental in revealing the polarized distribution of CELSR1 at hair cell to supporting cell (HC:SC) junctions in vestibular tissues .

• Western Blotting: Detecting CELSR1 protein expression levels and assessing protein integrity. This is particularly useful when evaluating deletion constructs or testing expression of CELSR1 variants .

• Immunoprecipitation: Isolating CELSR1 protein complexes to identify binding partners and study protein-protein interactions that mediate its functions in planar cell polarity .

• Immunohistochemistry: Examining the tissue distribution patterns of CELSR1 in fixed tissue sections, particularly useful for developmental studies .

• ELISA: Quantitative measurement of CELSR1 levels in biological samples .

When designing experiments, researchers should validate antibodies for their specific application, as performance can vary considerably between applications even for the same antibody .

What dilutions and incubation conditions are typically recommended for CELSR1 antibodies?

Optimal dilutions and incubation conditions for CELSR1 antibodies vary depending on the specific antibody and application. Based on the available data, the following general guidelines can be considered:

For immunofluorescence applications, successful protocols have used dilutions ranging from 1:200 (for guinea pig anti-Celsr1) to 1:400 (for rabbit anti-Celsr1 C-terminal antibodies) . For immunohistochemistry, dilutions typically range from 1:100 to 1:500 depending on the antibody sensitivity and tissue preparation .

Most commercially available CELSR1 antibodies are supplied in storage buffers containing PBS with 0.02% sodium azide and 50% glycerol at pH 7.3 . For long-term storage, antibodies should be kept at -20°C, where they remain stable for approximately one year after shipment . Small volume sizes (e.g., 20μl) may contain 0.1% BSA as a stabilizer . When working with these antibodies, avoid repeated freeze-thaw cycles and consider aliquoting for regular use.

How can researchers accurately visualize CELSR1's asymmetric distribution at cell boundaries?

Visualizing CELSR1's asymmetric distribution at cell boundaries requires meticulous experimental design and imaging techniques. Based on successful approaches, researchers should:

• Tissue Preparation: Utilize either whole-mount preparations or cryosections depending on the tissue of interest. For inner ear tissues, careful dissection of sensory epithelia followed by minimal fixation (typically 10-15 minutes in 4% paraformaldehyde) helps preserve protein localization while allowing antibody penetration .

• Antibody Selection: Choose antibodies recognizing specific domains of CELSR1. Antibodies against the cytoplasmic domain (such as those targeting amino acids 2885-2985) have successfully revealed asymmetric distributions in sensory epithelia . Co-staining with cell-type specific markers helps delineate boundaries between different cell populations.

• Confocal Microscopy: High-resolution confocal microscopy with z-stack acquisition is essential for accurately visualizing the three-dimensional distribution of CELSR1 at cell boundaries. For studying planar cell polarity, both en face (surface) views and cross-sectional views should be analyzed .

• Quantitative Analysis: To objectively assess asymmetric distribution, implement quantitative fluorescence intensity measurements along cell boundaries using appropriate imaging software. Plot intensity profiles around cell perimeters to demonstrate polarized distribution .

Research has shown that CELSR1 exhibits complex distribution patterns - in some tissues like semi-circular canal cristae, it shows clear polarized localization at specific boundaries, while in other regions it may surround individual hair cells . This heterogeneity highlights the importance of analyzing multiple samples and regions within tissues to accurately characterize CELSR1 distribution patterns.

What are the methodological approaches to investigating the functional domains of CELSR1?

Investigating the functional domains of CELSR1 requires strategic experimental approaches that dissect the protein's structure-function relationships. Based on published methodologies, researchers can employ the following strategies:

• Domain Deletion Constructs: Create targeted deletion constructs that remove specific functional modules of CELSR1. Research has successfully employed this approach by generating constructs lacking either the CADH1-8 module (ΔCADH1-8) or the CADH9-GAIN module (ΔCADH9-GAIN) . These constructs should be tagged with fluorescent reporters to confirm expression and localization.

• Expression Validation: Before functional analysis, validate expression of constructs using multiple approaches:

  • Fluorescence microscopy to confirm cellular localization

  • Western blotting to verify protein size and expression levels

  • Cell surface biotinylation to quantify membrane expression

• Functional Assays:

  • Cell Aggregation Assays: Mix differentially labeled cell populations expressing CELSR1 constructs and quantify the formation of multicellular aggregates. This directly tests homophilic adhesion capacity .

  • Cell-Cell Junction Enrichment Assays: Co-stain for CELSR1 constructs and junction markers like ZO-1 to assess localization to cell-cell contacts .

  • Signaling Activity Assays: Measure downstream signaling pathways affected by CELSR1 to determine which domains influence signaling versus adhesion functions .

These methodologies have revealed critical insights, such as the finding that the CADH1-8 module is essential for CELSR1-mediated cell-cell adhesion while neither the CADH1-8 nor CADH9-GAIN deletion affected signaling activity . This methodological framework allows researchers to dissect the multifunctional nature of CELSR1 domains.

How can researchers differentiate between the functions of CELSR1 versus other CELSR family members?

Differentiating between the functions of CELSR1 and other CELSR family members (CELSR2 and CELSR3) requires multifaceted experimental strategies that account for their overlapping yet distinct roles:

• Specific Antibody Selection: Use highly specific antibodies that do not cross-react between CELSR family members. Validate antibody specificity using tissues from knockout models or cells with CRISPR/Cas9-mediated deletion of individual CELSR genes . When working with antibodies, ensure they recognize species-specific epitopes unique to each CELSR protein.

• Genetic Approaches:

  • Generate and analyze single, double, and triple knockout models to identify unique versus redundant functions

  • Employ conditional knockout strategies in specific tissues to bypass embryonic lethality that may occur with global deletion

  • Use genetic rescue experiments where one CELSR family member is expressed in the background of another member's deletion

• Comparative Functional Analysis: Research has shown that Celsr1-3 differentially regulate neuronal migration, with Celsr1 specifically helping to specify migration direction . Design experiments that:

  • Compare phenotypes in cellular models where each CELSR is independently manipulated

  • Analyze the localization patterns of each CELSR in the same tissue context

  • Examine temporal expression patterns during development

• Domain-Specific Approaches: Create chimeric constructs that swap domains between CELSR family members to identify which regions confer functional specificity versus which domains have conserved functions across family members .

These approaches have revealed that while all CELSR proteins are involved in planar cell polarity, they have distinct roles - for instance, CELSR1 has been specifically implicated in vestibular hair cell polarity and directional neuronal migration , functions that are not fully compensated by CELSR2 or CELSR3.

What are the technical challenges in studying CELSR1 protein interactions and how can they be overcome?

Studying CELSR1 protein interactions presents several technical challenges due to its large size (329 kDa), complex domain structure, and membrane localization. Based on research approaches, these challenges can be addressed through:

• Protein Extraction and Solubilization:

  • CELSR1's seven-pass transmembrane structure makes it difficult to extract in native conformation

  • Utilize specialized lysis buffers containing mild detergents (e.g., 1% Triton X-100 or CHAPS) that preserve protein-protein interactions while solubilizing membrane proteins

  • Consider crosslinking approaches prior to cell lysis to stabilize transient interactions

• Co-Immunoprecipitation Optimization:

  • Use antibodies against specific domains of CELSR1 (e.g., cytoplasmic C-terminal region) for immunoprecipitation

  • Include appropriate controls including IgG controls and CELSR1-null samples

  • Consider tandem affinity purification approaches with tagged CELSR1 constructs to increase specificity and reduce background

• Domain-Specific Interaction Analysis:

  • Instead of working with full-length protein, study interactions of individual domains expressed as soluble fragments

  • For extracellular interactions, use secreted Fc-fusion proteins containing specific cadherin domains

  • For cytoplasmic interactions, use GST-pulldown or yeast two-hybrid approaches with the intracellular domain

• Live-Cell Imaging Approaches:

  • Employ FRET (Förster Resonance Energy Transfer) or BiFC (Bimolecular Fluorescence Complementation) methods to visualize protein interactions in living cells

  • Split-GFP approaches can be particularly valuable for studying membrane protein interactions

• Proximity Labeling Methods:

  • Implement BioID or APEX2 proximity labeling, where CELSR1 is fused to a biotin ligase that biotinylates proteins in close proximity

  • These approaches can identify the neighborhood of proteins surrounding CELSR1 at cell junctions

By combining these approaches, researchers can overcome the inherent difficulties in studying interactions of this large transmembrane protein while generating reliable data on its binding partners and functional complexes.

What are the critical controls needed when validating CELSR1 antibody specificity?

Validating CELSR1 antibody specificity is essential for generating reliable research data. Based on best practices, researchers should implement the following critical controls:

• Genetic Validation:

  • Test antibodies on tissues or cells from CELSR1 knockout/knockdown models

  • If knockout models are unavailable, use siRNA or CRISPR/Cas9 to reduce CELSR1 expression

  • Observe corresponding reduction or elimination of signal with specific antibodies

• Peptide Competition Assays:

  • Pre-incubate the antibody with the immunizing peptide prior to application

  • A specific antibody will show reduced or eliminated signal when blocked with its target epitope

  • Use non-relevant peptides as negative controls to confirm specificity of blocking

• Multiple Antibody Validation:

  • Compare staining patterns using antibodies targeting different epitopes of CELSR1

  • Consistent localization patterns with antibodies against different domains (e.g., extracellular versus cytoplasmic) provide strong evidence for specificity

  • For example, compare results between guinea pig anti-Celsr1 and rabbit anti-Celsr1 C-terminal antibodies

• Correlation with Tagged Constructs:

  • Express epitope-tagged CELSR1 (e.g., GFP-CELSR1) and compare antibody staining with tag detection

  • Co-localization between antibody signal and tag signal confirms specificity

• Western Blot Analysis:

  • Verify that the antibody detects a protein of the expected molecular weight (329 kDa)

  • Check for absence of non-specific bands or cross-reactivity with related proteins

  • Include positive controls (tissues known to express CELSR1) and negative controls

• Cross-Species Validation:

  • If the antibody claims reactivity with multiple species, test specificity across these species

  • Confirm that staining patterns align with known expression patterns in each species

Implementing these validation approaches ensures that experimental observations genuinely reflect CELSR1 biology rather than artifacts of non-specific antibody binding.

What cell models are most appropriate for studying CELSR1 function?

Selecting appropriate cell models is crucial for studying CELSR1 function effectively. Based on successful research approaches, consider the following model systems:

• HEK293T Cells: These cells have been effectively used for:

  • Expressing wild-type and mutant CELSR1 constructs

  • Cell aggregation assays to study CELSR1-mediated adhesion

  • Cell-cell junction enrichment assays with co-staining for junction markers like ZO-1

  HEK293T cells offer advantages of high transfection efficiency and low endogenous CELSR1 expression, providing a clean background for functional studies.

• Epithelial Cell Lines: Models such as MDCK or Caco-2 cells are valuable for:

  • Studying CELSR1's role in epithelial polarity

  • Examining localization to specific membrane domains

  • Investigating interactions with other polarity proteins

• Primary Cells:

  • Inner ear sensory epithelial cells for studying CELSR1's role in hair cell polarity

  • Neuronal cultures for examining CELSR1's function in neuronal migration and axon guidance

  • These primary systems maintain physiological expression levels and native interaction partners

• Organoid Models:

  • Inner ear organoids or cerebral organoids can recapitulate developmental contexts

  • Allow for studying CELSR1 function in three-dimensional tissue architecture

  • More closely mimic the in vivo environment compared to monolayer cultures

The choice of model system should be guided by the specific aspect of CELSR1 biology under investigation. For basic mechanistic studies of protein domains, heterologous expression systems like HEK293T cells are appropriate . For developmental and tissue-specific functions, primary cells or organoid models that better represent the native context are preferable .

What techniques are most effective for visualizing CELSR1 localization in developing tissues?

Visualizing CELSR1 localization in developing tissues requires specialized techniques that preserve tissue architecture while enabling high-resolution imaging. Based on successful approaches, researchers should consider:

• Tissue Preparation Methods:

  • For embryonic tissues, short fixation times (10-15 minutes) with 4% paraformaldehyde preserve antigenicity

  • For inner ear tissues, careful microdissection to expose sensory epithelia followed by gentle fixation is optimal

  • Consider vibratome sectioning for thick tissues to maintain 3D structure

• Immunofluorescence Approaches:

  • Use high-affinity, validated antibodies against specific domains of CELSR1

  • Implement antigen retrieval methods if necessary to expose epitopes

  • For membrane proteins like CELSR1, mild permeabilization conditions maintain native localization

  • Counter-stain with markers of specific cell types to provide cellular context

• Imaging Techniques:

  • Confocal microscopy with optical sectioning to visualize CELSR1 at specific cellular interfaces

  • Super-resolution microscopy (STED, STORM, or SIM) for nanoscale localization at cell junctions

  • Two-photon microscopy for deeper tissue imaging with reduced photobleaching

  • Light-sheet microscopy for rapid imaging of larger tissue volumes with reduced phototoxicity

• Analysis Approaches:

  • 3D reconstruction of z-stacks to visualize complex tissue architecture

  • Quantitative analysis of polarized protein distribution using line scan analysis

  • Co-localization analysis with junction markers and polarity proteins

These techniques have revealed important insights, such as CELSR1's asymmetric distribution at boundaries between hair cells and supporting cells in the developing vestibular system and its polarized localization in semi-circular canal cristae . The choice of specific approaches should be tailored to the developmental stage and tissue type under investigation.

How should researchers quantitatively analyze CELSR1 polarized distribution in tissues?

Quantitative analysis of CELSR1's polarized distribution requires rigorous methodological approaches to objectively measure asymmetry. Based on established research practices, implement these analytical strategies:

• Standardized Image Acquisition:

  • Use consistent microscope settings across all samples

  • Acquire images at optimal resolution to resolve cell boundaries

  • Collect z-stacks to capture the full three-dimensional distribution

  • Include internal controls within each image for normalization

• Boundary Intensity Analysis:

  • Trace cell boundaries manually or using semi-automated methods

  • Measure fluorescence intensity along the entire perimeter of cells

  • Plot intensity as a function of position to visualize polarity

  • Normalize to membrane markers to account for membrane density variations

• Polarity Vectors:

  • Assign vectors to represent the direction and magnitude of CELSR1 asymmetry

  • Calculate the vector sum for individual cells or tissue regions

  • Generate rose diagrams or circular histograms to visualize directional bias

  • Perform statistical analysis using circular statistics to quantify consistency of orientation

• Quantitative Metrics:

  Measurement	Calculation Method	Interpretation
  Polarity Index	(Imax-Imin)/(Imax+Imin)	Values close to 1 indicate strong polarity
  Angular Deviation	Standard deviation of polarity angles	Lower values indicate consistent orientation
  Enrichment Ratio	Intensity at specific boundary/average boundary intensity	Values >1 indicate enrichment

• Comparative Analysis:

  • Compare CELSR1 distribution with other polarity proteins

  • Analyze how distribution changes across developmental stages

  • Quantify differences between wild-type and mutant/treated samples

These quantitative approaches have revealed that CELSR1 maintains distinct polarized distributions at supporting cell-to-supporting cell junctions throughout vestibular cristae, while showing variable patterns at hair cell-to-supporting cell boundaries . Such nuanced analysis helps distinguish between global polarity defects and boundary-specific alterations in protein localization.

What are the most common technical challenges when working with CELSR1 antibodies?

Researchers frequently encounter several technical challenges when working with CELSR1 antibodies. Based on research experience, here are the most common issues and their solutions:

• High Background Signal:

  • Cause: Non-specific antibody binding or autofluorescence

  • Solution: Implement more stringent blocking (5% BSA or 10% serum), increase washing steps, and test different antibody dilutions. For tissues with high autofluorescence, consider Sudan Black B treatment or use of TrueBlack® quenching reagents .

• Poor Signal Detection:

  • Cause: Low antibody affinity, limited epitope accessibility, or low CELSR1 expression

  • Solution: Optimize antigen retrieval methods (heat-induced or enzymatic), test different fixation protocols, and increase antibody incubation time (overnight at 4°C). For tissues with low expression, consider tyramide signal amplification systems .

• Inconsistent Staining Patterns:

  • Cause: Variability in sample preparation or antibody batch differences

  • Solution: Standardize fixation duration and conditions, process all comparative samples simultaneously, and validate each new antibody lot against previous results .

• Issues with Western Blot Detection:

  • Cause: CELSR1's large size (329 kDa) makes transfer and detection challenging

  • Solution: Use low percentage gels (4-6%), extend transfer time with reduced voltage, implement wet transfer systems for large proteins, and use specialized transfer buffers containing SDS for large proteins .

• Cross-Reactivity with Other CELSR Family Members:

  • Cause: Sequence homology between CELSR1, CELSR2, and CELSR3

  • Solution: Select antibodies targeting unique epitopes, validate specificity using knockout controls, and confirm results with multiple antibodies targeting different regions of CELSR1 .

• Poor Reproducibility Between Antibodies:

  • Cause: Different epitope recognition affecting detection of protein subpopulations

  • Solution: Characterize each antibody's specific binding properties, use multiple antibodies recognizing different domains, and interpret results in the context of each antibody's limitations .

By anticipating these challenges and implementing appropriate solutions, researchers can generate more reliable and reproducible results when working with CELSR1 antibodies.

How can researchers optimize immunoprecipitation protocols for CELSR1?

Optimizing immunoprecipitation (IP) protocols for CELSR1 requires addressing the challenges associated with this large, multi-pass transmembrane protein. Based on successful approaches, implement these strategies:

• Lysis Buffer Optimization:

  • Use mild detergents that solubilize membranes while preserving protein interactions

  • Test different detergent combinations (e.g., 1% Triton X-100, 0.5% NP-40, or 0.5% digitonin)

  • Include protease inhibitors to prevent degradation

  • For protein complexes, consider including phosphatase inhibitors to maintain phosphorylation states

• Antibody Selection and Application:

  • Choose antibodies with high affinity for native CELSR1 conformation

  • Determine optimal antibody-to-lysate ratios through titration experiments

  • Pre-clear lysates with protein A/G beads to reduce non-specific binding

  • Consider covalently coupling antibodies to beads to prevent antibody contamination in eluates

• IP Conditions:

  • Extend incubation times (overnight at 4°C) to maximize protein capture

  • Use gentle rotation rather than shaking to preserve protein complexes

  • Optimize washing stringency - multiple gentle washes with decreasing detergent concentrations

  • For weak interactions, consider chemical crosslinking before lysis

• Elution Strategies:

  • For standard IP: Use low pH glycine buffer with immediate neutralization

  • For mass spectrometry analysis: Consider on-bead digestion to avoid contamination

  • For native complex isolation: Use competitive elution with immunizing peptide

• Controls and Validation:

  • Include negative controls (non-specific IgG) processed identically

  • Use lysates from CELSR1-depleted cells as additional negative controls

  • Validate IP efficiency by immunoblotting input, unbound, and eluted fractions

  • Confirm specific co-immunoprecipitation of known interacting partners

These optimization strategies have been successfully applied in studies examining CELSR1 interactions with other proteins involved in planar cell polarity pathways. By systematically testing and refining these parameters, researchers can develop robust IP protocols that yield specific and reproducible results with CELSR1.

What emerging technologies might advance CELSR1 research?

Emerging technologies offer promising avenues to overcome current limitations in CELSR1 research. Based on technological trends and research needs, these approaches may significantly advance our understanding:

• CRISPR-based Technologies:

  • Precision genome editing to create knockin models with endogenous fluorescent tags

  • CRISPRi/CRISPRa systems for temporal control of CELSR1 expression

  • Base editing for introducing specific point mutations linked to human disorders

  • CRISPR screens to identify genetic modifiers of CELSR1 function

• Advanced Imaging Technologies:

  • Lattice light-sheet microscopy for long-term imaging of CELSR1 dynamics in developing tissues

  • Super-resolution techniques (STORM, PALM, STED) to visualize nanoscale organization at cell junctions

  • Expansion microscopy to physically enlarge specimens for improved resolution of CELSR1 localization

  • Correlative light and electron microscopy (CLEM) to relate CELSR1 distribution to ultrastructural features

• Proximity Labeling Approaches:

  • TurboID or miniTurbo fusion proteins for rapid biotin labeling of CELSR1-proximal proteins

  • Split-TurboID systems to identify context-specific interaction partners

  • These methods can map the dynamic protein neighborhood of CELSR1 at specific developmental stages

• Single-Cell Technologies:

  • Single-cell transcriptomics to correlate CELSR1 expression with cell fate decisions

  • Single-cell proteomics to measure CELSR1 protein levels and modifications

  • Spatial transcriptomics to map CELSR1 expression in tissue context

• Structural Biology Approaches:

  • Cryo-electron microscopy of CELSR1 complexes to determine interaction mechanisms

  • AlphaFold2 and other AI-based structure prediction tools to model domain interactions

  • Hydrogen-deuterium exchange mass spectrometry to map conformational changes upon ligand binding

• Organoid and Tissue Engineering:

  • Advanced organoid systems that better recapitulate tissue architecture for studying CELSR1 in development

  • Bioengineered tissues with controlled geometric constraints to study mechanical aspects of CELSR1 function

  • Microfluidic organ-on-chip platforms for real-time manipulation and observation of CELSR1 dynamics

These technologies promise to provide unprecedented insights into CELSR1's function in development, tissue homeostasis, and disease contexts, potentially leading to new therapeutic strategies for disorders linked to CELSR1 dysfunction.

What are the critical unanswered questions in CELSR1 biology that antibody-based approaches might help resolve?

Several critical unanswered questions in CELSR1 biology could be addressed through strategic application of antibody-based approaches:

• Post-translational Modification Landscape:

  • Development of modification-specific antibodies (phospho-CELSR1, glycosylated-CELSR1) could reveal how CELSR1 function is regulated through PTMs

  • These tools would enable mapping of modification sites during development and in response to signaling

  • Combining IP with mass spectrometry using these antibodies could identify novel regulatory modifications

• Domain-Specific Functions in Different Tissues:

  • Antibodies recognizing specific CELSR1 domains could help determine if domain exposure varies across tissues

  • Epitope masking studies using domain-specific antibodies might reveal conformational changes during activation

  • These approaches could explain tissue-specific functions of CELSR1 in development and disease

• Dynamic Protein Complex Assembly:

  • Proximity ligation assays using CELSR1 antibodies with antibodies against suspected partners could map interaction networks in situ

  • Temporal analysis of these interactions during development would reveal when and where specific complexes form

  • This could explain how CELSR1 participates in distinct processes like cell adhesion and directional migration

• Internalization and Trafficking Mechanisms:

  • Antibodies against extracellular epitopes combined with endocytosis assays could track CELSR1 internalization

  • Surface biotinylation combined with anti-CELSR1 immunoprecipitation could quantify membrane versus intracellular pools

  • These approaches might resolve how CELSR1 trafficking regulates its asymmetric distribution

• Cleavage Products and Their Functions:

  • Antibodies recognizing specific regions could identify potential proteolytic processing of CELSR1

  • Domain-specific antibodies might detect previously unrecognized cleavage products with distinct functions

  • This could reveal whether CELSR1 undergoes processing similar to other adhesion GPCRs

• Resolution of Contradictory Findings:

  • In some tissues, CELSR1 shows clear polarized distribution, while in others it appears to surround individual cells

  • Carefully designed antibody studies could resolve whether these differences reflect true biological variation or technical artifacts

  • This might explain context-dependent functions of CELSR1 in different tissues

Addressing these questions through antibody-based approaches would significantly advance our understanding of CELSR1's multifaceted roles in development and disease, potentially identifying new therapeutic targets for conditions involving planar cell polarity defects.