PCM1 Antibody

What is PCM1 and why is it important in cellular research?

PCM1 (Pericentriolar Material 1) is a large protein (228.5 kDa) that functions as a key component of centriolar satellites, which are electron-dense granules scattered around centrosomes. This protein plays a critical role in centrosome assembly and function by correctly localizing several centrosomal proteins and anchoring microtubules to the centrosome . Research interest in PCM1 stems from its fundamental role in cell division and its association with various malignancies. Chromosomal aberrations involving the PCM1 gene have been linked to papillary thyroid carcinomas and hematological disorders, including atypical chronic myeloid leukemia and T-cell lymphoma . Multiple transcript variants encoding different isoforms have been identified, suggesting complex regulatory mechanisms. For researchers, PCM1 antibodies provide a means to study centrosome biology, cell cycle regulation, and disease mechanisms involving centrosomal abnormalities.

What applications are PCM1 antibodies validated for?

PCM1 antibodies are validated for multiple experimental applications, providing researchers with versatile tools for different research questions. The primary validated applications include:

• Western Blot (WB): For detection of PCM1 protein in cell and tissue lysates

• Immunohistochemistry (IHC): For visualization of PCM1 in tissue sections, particularly paraffin-embedded samples

• Immunofluorescence (IF): For subcellular localization studies

• Immunoprecipitation (IP): For protein-protein interaction studies

• Immunocytochemistry (ICC): For cellular localization in cultured cells

• Flow Cytometry (FCM): For quantitative analysis of PCM1 in cell populations

• ELISA: For quantitative measurement of PCM1 in samples

Each application requires specific validation parameters and optimization steps. For example, Western blot detection typically reveals bands at approximately 280 kDa (though the expected size is 229 kDa), and this discrepancy is likely due to post-translational modifications or the formation of protein complexes affecting migration patterns .

What are the different types of PCM1 antibodies available and how should I choose?

Researchers have several options when selecting PCM1 antibodies, each with distinct characteristics suited to different experimental purposes:

When selecting an antibody, consider:

• The species reactivity required (human, mouse, rat)

• The specific application (some antibodies perform better in certain applications)

• Whether you need a conjugated version (biotin, FITC, HRP, Alexa dyes)

• The region of PCM1 you wish to detect (some antibodies target specific domains)

For crucial experiments, validating the antibody in your specific experimental context is essential, regardless of supplier validation data.

How can I optimize PCM1 antibody usage for co-localization studies with other centrosomal proteins?

Co-localization studies of PCM1 with other centrosomal proteins require careful optimization to generate reliable data. Based on established protocols:

For immunofluorescence co-localization:

For reproducible results, process all experimental conditions in parallel and include appropriate controls for antibody specificity and fluorophore bleed-through.

What are the technical challenges in detecting PCM1 by Western blot, and how can they be addressed?

Detection of PCM1 by Western blot presents several technical challenges due to its high molecular weight (228.5 kDa) and complex structure. Researchers often observe a band around 280 kDa rather than the expected 229 kDa, indicating possible post-translational modifications . Key challenges and solutions include:

• Protein extraction efficiency:

  • Challenge: Complete extraction of high molecular weight proteins from centriolar satellites.

  • Solution: Use RIPA buffer supplemented with 1% SDS and mechanical disruption (sonication with 3-5 brief pulses). Avoid excessive sonication which can degrade PCM1.

• Gel electrophoresis optimization:

  • Challenge: Poor resolution and transfer of high molecular weight proteins.

  • Solution: Use 6-8% polyacrylamide gels rather than standard 10-12% gels. Extend running time at lower voltage (80V) to improve separation.

• Transfer efficiency:

  • Challenge: Incomplete transfer of large proteins to membrane.

  • Solution: Employ wet transfer systems with Towbin buffer containing 0.05-0.1% SDS and 10% methanol at 30V overnight at 4°C. Consider using PVDF membranes (0.45 μm pore size) which may provide better binding capacity than nitrocellulose for large proteins.

• Antibody concentration and incubation:

  • Challenge: Weak signal or high background.

  • Solution: Optimize primary antibody concentration (typically 1:500 to 1:2000) and incubate overnight at 4°C. Extended washing (5 x 10 minutes) can significantly reduce background while maintaining specific signal.

• Multiple band interpretation:

  • Challenge: Multiple bands may represent isoforms, degradation products, or non-specific binding.

  • Solution: Include positive controls (e.g., HeLa or 293T cell lysates) alongside experimental samples. Consider using isoform-specific antibodies when investigating particular variants .

A panel of loading controls is recommended, as traditional controls like β-actin (42 kDa) may not accurately reflect loading of high molecular weight proteins.

How can PCM1 antibodies be used to study PCM1's role in genetic disorders and cancer?

PCM1 has been implicated in various genetic disorders and cancers, particularly through chromosomal rearrangements. PCM1 antibodies provide valuable tools for investigating these associations through multiple approaches:

• Diagnostic immunohistochemistry (IHC):
  PCM1 antibodies can detect abnormal PCM1 expression patterns in clinical samples. In papillary thyroid carcinomas associated with RET/PCM1 rearrangements, IHC can reveal altered subcellular localization. Optimize antigen retrieval using EDTA buffer (pH 8.0) for paraffin-embedded sections and titrate antibody concentration (typically 1:50 to 1:200) for cancer tissue microarrays .

• Fusion protein detection:
  PCM1-JAK2 and PCM1-RET fusion proteins, associated with leukemias and thyroid cancers respectively, can be detected using Western blot with PCM1 antibodies directed against N-terminal epitopes. This approach allows visualization of fusion proteins with altered molecular weights compared to wild-type PCM1.

• Cell model systems:
  For functional studies, combine PCM1 antibodies with:

  • siRNA/shRNA knockdown validation

  • CRISPR-engineered cell lines

  • Ectopic expression of fusion constructs

  Monitor centrosome structure and function via immunofluorescence co-staining with γ-tubulin or centrin markers to correlate PCM1 alterations with centrosomal abnormalities.

• Proximity ligation assay (PLA):
  This technique can detect protein-protein interactions at endogenous levels. Using PCM1 antibodies in combination with antibodies against suspected interaction partners can reveal altered interaction networks in disease states.

• Chromatin immunoprecipitation (ChIP) studies:
  For PCM1 fusion proteins with transcription factors, ChIP using PCM1 antibodies can identify altered DNA binding and transcriptional regulation.

These methodologies enable researchers to connect PCM1 aberrations to phenotypic outcomes and molecular mechanisms in genetic disorders and cancer models .

What are the best practices for immunoprecipitation using PCM1 antibodies?

Successful immunoprecipitation (IP) of PCM1 requires specific technical considerations due to its large size and complex interactions with other centrosomal proteins. The following protocol incorporates key optimization steps:

• Lysis buffer selection:
  For PCM1 IP, use a gentle lysis buffer to preserve protein-protein interactions:

  • 25 mM Tris-HCl (pH 7.4)

  • 150 mM NaCl

  • 1% NP-40 or 0.5% Triton X-100

  • 1 mM EDTA

  • 5% glycerol

  • Freshly added protease and phosphatase inhibitor cocktails

• Pre-clearing optimization:
  Pre-clear lysates with 20 μL of Protein A/G beads per 1 mg of protein for 1 hour at 4°C to reduce non-specific binding. This step is particularly important for PCM1 due to its tendency to form large complexes that may bind non-specifically.

• Antibody selection and binding:

  • For PCM1 IP, use 2-5 μg of antibody per 500 μg of total protein

  • Incubate antibody with lysate overnight at 4°C with gentle rotation

  • G-6 clone antibodies have demonstrated high efficiency in IP applications with consistent results

• Bead incubation and washing:

  • Add 30-50 μL of Protein A/G beads per sample and incubate for 2-3 hours at 4°C

  • Perform stringent washing: 4 washes with lysis buffer followed by 2 washes with PBS containing 0.1% Tween-20

  • Perform a final wash with PBS only to remove detergent

• Elution strategies:

  • Gentle elution: 0.1 M glycine (pH 2.5-3.0) for 10 minutes at room temperature, followed by neutralization with 1M Tris (pH 8.0)

  • Denaturing elution: 1X Laemmli buffer with 5% β-mercaptoethanol, heated at 70°C (not 95°C) for 10 minutes to minimize aggregation of high molecular weight PCM1

• Controls:

  • IgG control: Use species-matched IgG at the same concentration

  • Input control: Load 5-10% of pre-IP lysate

  • Validation: Confirm IP success with Western blot using a PCM1 antibody recognizing a different epitope from the IP antibody

For co-immunoprecipitation studies investigating PCM1 binding partners, cross-linking with 1% formaldehyde prior to lysis may help preserve transient interactions.

How should PCM1 antibodies be validated to ensure specificity in different experimental systems?

Comprehensive validation of PCM1 antibodies is essential for generating reliable experimental data. The following multi-step validation approach addresses potential pitfalls:

• Western blot validation:

  • Confirm detection of a band at ~228-280 kDa in positive control cell lines (HeLa, 293T, HT-1080)

  • Include negative controls via PCM1 knockdown (siRNA/shRNA) or knockout (CRISPR/Cas9) cells

  • Verify band disappearance with pre-adsorption using immunizing peptide

  • Test antibody specificity across species if cross-reactivity is claimed (human, mouse, rat)

• Immunofluorescence validation:

  • Compare staining pattern with established PCM1 localization (pericentriolar granules surrounding γ-tubulin-positive centrosomes)

  • Perform co-localization with alternative PCM1 antibodies recognizing different epitopes

  • Confirm signal reduction in PCM1-depleted cells

  • Test specificity for different PCM1 isoforms through rescue experiments with isoform-specific constructs

• Mass spectrometry validation:

  • Perform immunoprecipitation followed by LC-MS/MS analysis

  • Confirm PCM1 as one of the top hits in the precipitated complex

  • Evaluate enrichment of known PCM1-interacting proteins (e.g., BBS4, CEP290)

• Cross-platform validation:
  Create a validation matrix across multiple techniques:

  Validation Method	Primary Goal	Secondary Confirmation
  Western Blot	Confirm molecular weight	Test multiple cell lines/tissues
  Immunofluorescence	Verify subcellular localization	Co-localization with other centrosomal markers
  Immunoprecipitation	Confirm ability to pull down PCM1	Analyze co-precipitating proteins
  Genetic models	Test specificity via knockdown/knockout	Rescue experiments with tagged constructs
  Peptide blocking	Confirm epitope specificity	Titrate blocking peptide concentrations

• Application-specific validation:

  • For IHC applications, include tissue microarrays with known PCM1 expression patterns

  • For flow cytometry, compare cell surface vs. intracellular staining protocols

  • For super-resolution microscopy, validate resolution-dependent localization patterns

Complete validation data should be documented and considered when interpreting experimental results, especially when comparing studies using different antibodies or detection methods.

What are the optimal fixation and permeabilization conditions for PCM1 antibody staining in immunocytochemistry?

PCM1's unique localization to centriolar satellites requires careful optimization of fixation and permeabilization conditions for accurate visualization. Different experimental questions may require tailored protocols:

The optimal protocol should be empirically determined for each cell type and PCM1 antibody combination. For super-resolution microscopy applications, additional optimization of fixation timing and antibody concentration is often necessary to achieve the best signal-to-noise ratio.

Why does PCM1 sometimes appear at a different molecular weight than expected in Western blots?

PCM1 often appears at approximately 280 kDa in Western blots despite its calculated molecular weight of 228.5 kDa . This discrepancy can be attributed to several biological and technical factors:

• Post-translational modifications (PTMs):
  PCM1 undergoes extensive phosphorylation, particularly during cell cycle progression. Multiple kinases including PLK1 and CDK1 phosphorylate PCM1, adding negative charges that can reduce electrophoretic mobility. Additionally, PCM1 contains predicted sites for other PTMs including SUMOylation and ubiquitination, which can significantly alter apparent molecular weight.

• Alternative splicing:
  Multiple transcript variants of PCM1 have been identified, potentially producing protein isoforms with different molecular weights. The presence of tissue-specific or cell-cycle-dependent alternative splicing can result in band shifts or multiple bands on Western blots.

• Technical factors affecting migration:

  • Incomplete protein denaturation: PCM1's large size and complex structure may resist complete denaturation, affecting migration

  • Salt concentration: High salt in samples can cause aberrant migration patterns

  • Acrylamide percentage: Lower percentage gels (6-8%) provide better resolution for high molecular weight proteins like PCM1

• Protein-protein interactions:
  Despite denaturing conditions, very stable protein complexes may occasionally remain intact during SDS-PAGE, resulting in apparent higher molecular weight bands.

• Interpretation guidelines:

  Observed Pattern	Likely Explanation	Verification Approach
  Single band at ~280 kDa	Normal PCM1 with PTMs	Compare across multiple cell types
  Multiple bands (280, 228, smaller bands)	Isoforms and/or degradation	Phosphatase treatment, isoform-specific antibodies
  Smeared appearance	Heterogeneous phosphorylation	Lambda phosphatase treatment
  Higher bands (>300 kDa)	Potential dimers or stable complexes	Stronger denaturation conditions
  Only smaller bands (<200 kDa)	Likely degradation	Fresh sample preparation, additional protease inhibitors

To confirm the identity of PCM1 bands, researchers should:

• Use multiple antibodies targeting different PCM1 epitopes

• Include controls with PCM1 knockdown or overexpression

• Consider phosphatase treatment to eliminate migration shifts due to phosphorylation

• Use mass spectrometry for definitive identification of unexpected bands

How can I distinguish between specific and non-specific staining when using PCM1 antibodies in immunofluorescence?

Distinguishing specific PCM1 staining from artifacts in immunofluorescence requires systematic controls and careful interpretation of staining patterns:

• Characteristic PCM1 staining pattern:
  Authentic PCM1 staining typically appears as discrete punctate structures clustered around centrosomes. These pericentriolar granules (centriolar satellites) should:

  • Concentrate around the centrosome (co-stain with γ-tubulin or centrin)

  • Show dynamic redistribution during mitosis

  • Disperse upon microtubule depolymerization (nocodazole treatment)

  • Demonstrate characteristic density and size distribution (~70-100 nm granules visualized as diffraction-limited spots by conventional microscopy)

• Essential controls to include:

  • Genetic controls: PCM1 knockdown/knockout cells should show significant reduction in specific signal

  • Antibody controls: Pre-immune serum or isotype-matched IgG at equivalent concentration

  • Peptide competition: Pre-incubation of antibody with immunizing peptide should abolish specific staining

  • Secondary-only control: Omit primary antibody to assess secondary antibody specificity

  • Cross-validation: Compare staining pattern with alternative PCM1 antibodies targeting different epitopes

• Distinguishing features of non-specific staining:

  • Non-specific nuclear staining: Often uniform rather than punctate

  • Golgi-like staining: PCM1 can sometimes appear to localize to Golgi; confirm with Golgi markers

  • Edge artifacts: Increased staining at cell periphery regardless of cell type

  • Fixation artifacts: Signal that varies dramatically with different fixation methods

• Cell-type considerations:
  PCM1 expression and centriolar satellite organization vary across cell types:

  • Epithelial cells: Well-organized satellites around centrosomes

  • Neurons: PCM1 may localize to dendrites and neuronal projections

  • Ciliated cells: PCM1 localizes near basal bodies during ciliogenesis

  • Mitotic cells: PCM1 redistributes during mitosis, with satellites dissolving in metaphase

• Quantitative assessment:
  Implement quantitative approaches to distinguish specific from non-specific signals:

  • Signal-to-background ratio: Measure intensity of pericentriolar signal vs. cytoplasmic background

  • Colocalization coefficients: Calculate Pearson's or Mander's coefficients with known centrosomal markers

  • Intensity profiles: Generate line scans through centrosomes to visualize satellite distribution

  • 3D analysis: Z-stack acquisition to confirm 3D organization of satellites around centrosomes

For borderline cases, consider dual validation with live-cell imaging using fluorescently tagged PCM1 to confirm localization patterns observed with antibody staining.

What are common causes of inconsistent PCM1 antibody performance across experiments, and how can they be addressed?

Inconsistent PCM1 antibody performance can significantly impact experimental reproducibility. Understanding and addressing these variables is critical for reliable results:

• Antibody-related variables:

  • Lot-to-lot variability: Document lot numbers and test new lots against previous ones

  • Antibody degradation: Aliquot antibodies to minimize freeze-thaw cycles; store according to manufacturer recommendations

  • Concentration shifts: Periodically reconfirm optimal working concentration via titration experiments

• Cell and tissue sample variables:

  • Cell cycle effects: PCM1 localization and modification state changes throughout the cell cycle; synchronize cells when possible

  • Cell density effects: Confluence levels affect centrosome organization; standardize seeding density

  • Passage number: High passage cells may show altered centrosome structure; use cells within defined passage ranges

  • Sample preparation timing: PCM1 is sensitive to degradation; minimize time between sample collection and processing

• Protocol inconsistencies:
  Common technical variables affecting PCM1 detection include:

  Variable	Impact on PCM1 Detection	Standardization Approach
  Fixation timing	Overfixation masks epitopes	Standardize to exact minutes (e.g., precisely 10 min)
  Temperature fluctuations	Affects antibody binding kinetics	Use temperature-controlled incubators for all steps
  Buffer composition	pH and salt concentration affect epitope accessibility	Prepare buffers in bulk, aliquot and monitor pH
  Blocking effectiveness	Insufficient blocking increases background	Optimize blocking agent (BSA vs. serum) and time
  Washing stringency	Inadequate washing retains non-specific binding	Standardize washing duration, volume and agitation

• Technical approach for maximizing consistency:

  • Validation panel: Establish a panel of positive control samples with known PCM1 expression

  • Reference images: Maintain a library of reference images for comparison

  • Positive control inclusion: Process known positive samples alongside experimental samples

  • Batch processing: Process all experimental conditions simultaneously when possible

  • Internal controls: Include housekeeping protein controls processed identically

• Documentation practices:

  • Create detailed protocol worksheets recording all variables

  • Document reagent sources, catalog numbers, and lot numbers

  • Maintain equipment calibration logs (microscope settings, imager exposure times)

  • Record image acquisition parameters for all experiments

• Analytical approaches:

  • Implement quantitative analysis metrics rather than relying solely on visual assessment

  • Use automated image analysis algorithms to reduce subjective interpretation

  • Establish clear criteria for positive vs. negative staining before analyzing experimental samples

  • Consider blinding analysis to reduce confirmation bias

For critical experiments, replicate findings using alternative PCM1 antibodies or complementary techniques such as in situ hybridization or proximity ligation assays.

How are PCM1 antibodies being used to investigate centrosome biology and ciliopathies?

PCM1 antibodies have become essential tools for investigating the relationship between centriolar satellites, centrosome function, and ciliopathies (disorders of the primary cilium). Recent research applications include:

• Centriolar satellite dynamics and composition:
  PCM1 antibodies enable tracking of centriolar satellite formation, movement, and disassembly during the cell cycle. Immunofluorescence combined with live-cell imaging has revealed that PCM1-positive centriolar satellites undergo dynamic redistribution during cell cycle progression, with characteristic dispersion during mitosis and reconcentration in G1 phase. Researchers employ PCM1 antibodies in combination with super-resolution microscopy to map the precise architecture of centriolar satellites and their relationship to the centrosome.

• Ciliopathy protein interactions:
  PCM1 interacts with multiple proteins implicated in ciliopathies, including BBS4, CEP290, and OFD1. Research techniques utilizing PCM1 antibodies include:

  • Co-immunoprecipitation to identify novel PCM1-interacting proteins

  • Proximity ligation assays to verify interactions in situ

  • Sequential immunoprecipitation to isolate specific subcomplexes

  • Immunofluorescence co-localization to track spatial relationships during ciliogenesis

• Ciliogenesis regulation:
  PCM1 plays a critical role in primary cilium formation. Researchers use PCM1 antibodies to:

  • Track PCM1 redistribution during cilium initiation

  • Investigate PCM1's role in ciliary vesicle docking

  • Examine temporal sequences of protein recruitment during basal body maturation

  • Study the effect of ciliopathy mutations on PCM1 localization and function

• Cellular stress responses:
  Recent studies have implicated PCM1 in cellular stress responses, particularly those involving autophagy and proteostasis:

  • PCM1 antibodies are used to track its redistribution during autophagy induction

  • Co-staining with autophagy markers (LC3B, p62) reveals functional relationships

  • PCM1 antibodies help monitor satellite reorganization during heat shock and oxidative stress

• Disease model applications:
  PCM1 antibodies are critical for studying disease mechanisms in:

  • Patient-derived fibroblasts from ciliopathy patients

  • iPS-derived organoids modeling neurodevelopmental disorders

  • Animal models of retinal degeneration and kidney disease

  • Cancer cell lines with centrosomal amplification

As research continues to uncover PCM1's multifaceted roles, antibodies targeting specific PCM1 domains or modification states (phospho-specific antibodies) are becoming increasingly important for dissecting its precise functions in health and disease.

What methodological advances have improved the application of PCM1 antibodies in research?

Recent technological and methodological advances have significantly enhanced the utility and reliability of PCM1 antibodies in research applications:

• Advanced microscopy applications:
  Super-resolution microscopy has revolutionized the visualization of centriolar satellites:

  • Structured Illumination Microscopy (SIM) improves resolution to ~100 nm, allowing better visualization of individual satellites

  • Stochastic Optical Reconstruction Microscopy (STORM) achieves ~20 nm resolution, revealing internal organization of PCM1-containing structures

  • Expansion microscopy physically enlarges specimens, enabling conventional microscopes to resolve satellite substructures

  • Lattice light-sheet microscopy allows long-term live imaging of PCM1 dynamics with minimal phototoxicity

  These techniques require specific optimization of PCM1 antibody protocols, including adjusted concentration, specialized secondary antibodies, and modified sample preparation.

• Proximity-based interaction detection:
  Beyond traditional co-immunoprecipitation, newer techniques include:

  • BioID/TurboID: Proximity-dependent biotin labeling using PCM1 fusion proteins followed by streptavidin pulldown identifies neighborhood proteins

  • APEX2: Proximity-based labeling combined with PCM1 antibody immunoprecipitation reveals transient interactions

  • Proximity Ligation Assay (PLA): Detects proteins within 40 nm of PCM1 in fixed specimens

  • FRET-based interaction assays: Combined with PCM1 antibodies for endogenous protein validation

• Genome editing integration:
  CRISPR/Cas9 technology complementing PCM1 antibody approaches:

  • Endogenous tagging of PCM1 with small epitope tags (FLAG, HA, V5) for antibody detection without overexpression artifacts

  • Generation of validated PCM1 knockout cell lines as essential negative controls

  • PCM1 domain deletion mutants to map functional regions recognized by different antibodies

  • Introduction of patient-specific mutations to study pathogenic mechanisms

• Quantitative proteomics integration:
  Mass spectrometry approaches combined with PCM1 antibodies:

  • Multiplexed PCM1 interactome analysis using TMT or iTRAQ labeling

  • Cross-linking mass spectrometry to map PCM1 interaction surfaces

  • Phosphoproteomics to identify PCM1 modification sites during cell cycle progression

  • Absolute quantification of PCM1 levels in different cell types and conditions

• Automated imaging and analysis platforms:
  High-content screening approaches with PCM1 antibodies:

  • Automated image acquisition and analysis of PCM1 distribution following drug treatments

  • Machine learning algorithms for unbiased classification of PCM1 staining patterns

  • Quantitative analysis of PCM1 satellite number, size, and distribution

  • Correlative phenotype analysis linking PCM1 alterations to cellular functions

These methodological advances have elevated PCM1 antibody applications beyond descriptive observations to quantitative, systems-level analyses of centrosome biology and related pathologies.

How can PCM1 antibodies be used in studying neurodevelopmental disorders?

PCM1 has emerged as a significant factor in neurodevelopmental disorders, with genetic studies linking PCM1 variants to schizophrenia, autism spectrum disorders, and microcephaly. PCM1 antibodies provide critical tools for investigating these connections through various experimental approaches:

• Neurodevelopmental model systems:
  PCM1 antibodies enable investigation of neuronal centrosome and primary cilium function in:

  • Neural progenitor proliferation: Immunostaining reveals PCM1 distribution during symmetric vs. asymmetric divisions

  • Neuronal migration: PCM1 localization during cortical development correlates with migration defects

  • Neurite extension: PCM1-positive satellites distribute into developing axons and dendrites

  • Synaptogenesis: Potential roles for PCM1 in local protein synthesis at synaptic sites

• Brain organoid applications:
  Human iPSC-derived brain organoids provide 3D models for studying PCM1 function:

  • Temporal expression patterns during organoid development

  • Spatial organization in ventricular zone neural progenitors

  • Co-localization with microcephaly-associated proteins

  • Effects of patient-derived PCM1 mutations on organoid formation

• PCM1 in neurogenesis and differentiation:
  PCM1 antibodies reveal critical roles during neural differentiation:

  • Redistribution during neurogenic divisions of radial glia

  • Association with mother centriole during basal body formation

  • Co-localization with primary cilium during neural progenitor signaling

  • Satellite reorganization during neuronal maturation

• PCM1 in postmortem brain tissue:
  Analysis of human brain samples from patients with neurodevelopmental disorders:

  • Altered PCM1 expression or localization in affected brain regions

  • Changes in centriolar satellite organization in specific neuronal populations

  • Correlation between PCM1 distribution and neuronal morphology

  • Co-localization with disease-associated proteins

• Experimental approaches using PCM1 antibodies:

  Technique	Application in Neurodevelopmental Research	Key Considerations
  IHC in brain sections	Spatial distribution across brain regions	Requires optimized antigen retrieval for fixed tissue
  Primary neuron IF	Subcellular localization during differentiation	Combine with stage-specific neuronal markers
  Live imaging with PCM1 antibody fragments	Dynamics during neuronal migration	Requires specialized antibody delivery methods
  EM immunogold labeling	Ultrastructural localization in neuronal centrosomes	Needs carefully optimized fixation protocols
  Synaptosome fractionation + WB	PCM1 presence at synapses	Use multiple antibodies to confirm specificity

• PCM1 in neurodevelopmental signaling pathways:
  Recent studies implicate PCM1 in key signaling pathways relevant to neurodevelopment:

  • Sonic Hedgehog (Shh) signaling: PCM1 antibodies reveal relationships with ciliary signaling components

  • Wnt signaling: PCM1's interaction with Wnt pathway proteins affects neurogenesis

  • BMP/TGF-β pathways: PCM1's role in receptor trafficking impacts neural differentiation

These applications highlight how PCM1 antibodies serve as essential tools for connecting centrosomal dysfunction to neurodevelopmental pathologies, offering insights into both basic mechanisms and potential therapeutic approaches.