PTPN13 Antibody, FITC conjugated

What are the primary applications for FITC-conjugated PTPN13 antibodies in experimental research?

FITC-conjugated PTPN13 antibodies are primarily utilized in fluorescence-based applications, offering direct visualization of protein localization without requiring secondary antibodies. While unconjugated antibodies are commonly used in Western Blot (WB), Immunohistochemistry (IHC), and Immunofluorescence (IF) , FITC-conjugated versions excel in flow cytometry (FACS), immunofluorescence microscopy, and live-cell imaging experiments. The conjugation provides advantages for multiple labeling experiments, particularly when investigating PTPN13's role in cell junction formation and mesenchymal-to-epithelial transition . These antibodies enable researchers to track PTPN13 localization at intercellular junctions and desmosome formation in real-time visualization experiments . The fluorescent properties also facilitate quantitative analysis of PTPN13 expression levels in different cellular compartments, advancing our understanding of its subcellular distribution during cancer progression.

How should PTPN13 antibodies be stored to maintain optimal activity?

To maintain optimal activity of PTPN13 antibodies, researchers should follow specific storage protocols based on antibody formulation. Unconjugated antibodies are typically provided in 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 up to 12 months after shipment . For fluorophore-conjugated antibodies, including FITC conjugates, protection from light is essential to prevent photobleaching. After reconstitution, antibodies can be stored at 2-8°C under sterile conditions for approximately 1 month or at -20°C to -70°C for up to 6 months . Repeated freeze-thaw cycles should be strictly avoided by preparing small aliquots before freezing . For the 20μL size formats that contain 0.1% BSA, aliquoting is generally unnecessary for -20°C storage . Following these guidelines ensures the preservation of both binding specificity and fluorescence intensity, critical for reproducible experimental results.

What controls should be included when using FITC-conjugated PTPN13 antibodies in immunofluorescence studies?

When designing immunofluorescence experiments with FITC-conjugated PTPN13 antibodies, researchers must implement a comprehensive set of controls to ensure valid and interpretable results:

• Isotype control: Include a FITC-conjugated IgG from the same host species (rabbit for most PTPN13 antibodies) at the same concentration to assess non-specific binding.

• Negative tissue/cell controls: Use tissues or cell lines known to express minimal PTPN13, such as certain non-cancerous cell lines, to establish background fluorescence levels.

• Positive tissue/cell controls: Include samples with confirmed PTPN13 expression, such as MCF-7 cells or HeLa cell lysates , which have been validated in previous studies.

• Absorption controls: Pre-incubate the antibody with recombinant PTPN13 protein (such as the immunogen fusion protein) to confirm signal specificity.

• Secondary antibody-only control: For comparison with direct FITC conjugates, include samples with secondary antibody alone to assess non-specific fluorescence.

• Autofluorescence control: Examine unstained samples to identify any natural fluorescence from the tissue or fixation-induced artifacts.

• Subcellular marker co-localization: Include markers for cell junctions (given PTPN13's role in junction stabilization) to confirm proper localization patterns.

These controls collectively validate antibody specificity, optimize signal-to-noise ratios, and ensure accurate interpretation of PTPN13 localization and expression patterns.

How can I optimize antigen retrieval for PTPN13 detection in fixed tissue samples?

Optimizing antigen retrieval for PTPN13 detection requires careful consideration of tissue type and fixation method. Based on validated protocols, the following optimization strategy is recommended:

• Primary buffer selection: For formalin-fixed paraffin-embedded (FFPE) tissues, TE buffer at pH 9.0 is the preferred antigen retrieval solution for PTPN13 detection . Alternatively, citrate buffer at pH 6.0 can be used if TE buffer yields suboptimal results .

• Heat-induced epitope retrieval (HIER): For consistent results, perform HIER using either:

  • Microwave method: Heat samples in retrieval buffer to 95-98°C for 15-20 minutes, followed by cooling at room temperature for 20 minutes.

  • Pressure cooker method: Heat samples at maximum pressure for 3-5 minutes, followed by cooling.

• Tissue-specific considerations: For breast cancer and lung cancer tissues, which frequently express PTPN13 , additional optimization may be required:

  • Extend retrieval time to 25-30 minutes for densely fibrous breast tissues

  • Reduce retrieval time to 10-15 minutes for more fragile lung tissues

• Fixation impact assessment: Compare results from tissues fixed for different durations (6h, 12h, 24h) to determine optimal fixation conditions for PTPN13 epitope preservation.

• Antibody dilution optimization: After retrieval, test a range of antibody dilutions (1:50 to 1:500 for IHC applications) to determine optimal signal-to-noise ratio.

• Signal amplification: For tissues with low PTPN13 expression, implement tyramide signal amplification system after antigen retrieval to enhance detection sensitivity.

• Validation approach: Compare retrieval methods using both FITC-conjugated and unconjugated PTPN13 antibodies with appropriate visualization systems to confirm epitope accessibility.

This systematic approach ensures maximal epitope exposure while maintaining tissue morphology, critical for accurate PTPN13 localization and expression studies.

What is the recommended dilution range for FITC-conjugated PTPN13 antibodies across different applications?

The optimal dilution for FITC-conjugated PTPN13 antibodies varies by application, sample type, and detection system. While specific recommendations for FITC-conjugated versions must be determined empirically, guidance can be derived from established protocols for unconjugated PTPN13 antibodies:

For direct FITC conjugates, consider these additional factors:

• Fluorophore-to-protein ratio affects optimal dilution; higher ratios may require further dilution to prevent quenching

• FITC is more susceptible to photobleaching than other fluorophores, potentially requiring higher starting concentrations

• Autofluorescence in the FITC channel may necessitate further optimization, particularly in tissues with high natural fluorescence

For quantitative applications, a titration series is essential to determine the saturation point. Begin with the manufacturer's recommended range and perform sequential twofold dilutions, plotting mean fluorescence intensity against antibody concentration to identify the optimal dilution that provides maximum specific signal with minimal background . Always perform sample-dependent optimization, as noted in the literature, since cellular expression levels of PTPN13 vary significantly between different cancer cell lines and tissue types .

How can I effectively use PTPN13 antibodies to investigate its role in tumor suppression and cell junction stabilization?

To investigate PTPN13's tumor suppression and cell junction stabilization functions effectively, implement the following comprehensive research strategy:

• Comparative expression analysis: Use FITC-conjugated PTPN13 antibodies to quantify expression levels in normal vs. cancerous tissues through flow cytometry and immunofluorescence microscopy. Research has established that PTPN13 expression correlates with patient survival in multiple cancer types .

• Junction protein co-localization experiments: Design dual-labeling experiments using FITC-conjugated PTPN13 antibodies alongside markers for:

  • Adherens junctions (E-cadherin, β-catenin)

  • Desmosomes (desmoplakin, desmoglein)

  • Tight junctions (ZO-1, claudins)

  This approach will reveal PTPN13's specific association with junction complexes, supporting findings that PTPN13 stabilizes intercellular adhesion and promotes desmosome formation .

• Phosphorylation state analysis: Combine PTPN13 detection with phospho-specific antibodies targeting known PTPN13 substrates, including IRS-1 at Tyr612, which shows increased phosphorylation upon PTPN13 knockdown .

• Genetic modification approaches: In parallel with antibody-based detection:

  • Knockdown PTPN13 using siRNAs to assess effects on cell proliferation (demonstrated to increase growth rates in Schwann cells)

  • Compare wild-type cells with those expressing catalytically inactive PTPN13 mutants

  • Develop stable cell lines with modulated PTPN13 expression for long-term studies

• In vivo modeling: Apply FITC-conjugated PTPN13 antibodies for tumor imaging in:

  • HER2-overexpressing mice with wild-type PTPN13 versus mice lacking PTPN13 phosphatase activity

  • Xenograft models with PTPN13-overexpressing cancer cell lines to track mesenchymal-to-epithelial transition

• Functional assays: Correlate PTPN13 localization with:

  • Cell motility (wound healing assays)

  • Invasion capacity (Boyden chamber assays)

  • Intercellular adhesion strength (cell aggregation assays)

This integrated approach leverages fluorescent antibodies to connect PTPN13's molecular interactions with its functional roles in tumor suppression and cell adhesion.

What methods can be used to validate FITC-conjugated PTPN13 antibody specificity for critical research applications?

Validating the specificity of FITC-conjugated PTPN13 antibodies is crucial for generating reliable research data. A comprehensive validation strategy should include:

• Genetic validation approaches:

  • CRISPR/Cas9 knockout controls: Generate PTPN13-knockout cell lines to confirm absence of signal with the FITC-conjugated antibody

  • siRNA knockdown validation: Demonstrate reduced signal intensity proportional to knockdown efficiency, as established in previous PTPN13 studies

  • Overexpression systems: Compare signal in wild-type cells versus those overexpressing PTPN13 (both functional and catalytically inactive forms)

• Biochemical validation methods:

  • Immunoprecipitation followed by mass spectrometry: Confirm that the antibody captures PTPN13 protein (277 kDa)

  • Peptide competition assays: Pre-incubate antibody with immunizing peptide or recombinant PTPN13 fusion protein before application to samples

  • Western blot correlation: Confirm that IF/FACS signal intensity correlates with protein levels detected by validated Western blot antibodies, which should detect PTPN13 at approximately 260-277 kDa

• Cross-platform validation:

  • Orthogonal detection methods: Compare results with multiple PTPN13 antibodies recognizing different epitopes (AA 1-161, AA 250-500, AA 801-900, AA 1965-2173)

  • mRNA correlation: Correlate protein detection with PTPN13 mRNA levels via RT-qPCR

  • Reporter systems: Use cells expressing PTPN13-fluorescent protein fusions as reference standards

• Application-specific controls:

  • Isotype-matched negative controls: Use FITC-conjugated rabbit IgG at matching concentrations

  • Signal blocking with unconjugated antibody: Pre-incubate with excess unconjugated PTPN13 antibody

  • Multi-spectral imaging: Distinguish true signal from autofluorescence through spectral unmixing

• Cross-reactivity assessment:

  • Multi-species testing: Evaluate specificity across human, mouse, and rat samples based on known reactivity patterns

  • Phosphatase family cross-reactivity: Test against closely related phosphatases to confirm specificity

This comprehensive validation framework ensures that observed signals genuinely represent PTPN13 localization and expression, a critical consideration given its important role in cancer biology .

How can FITC-conjugated PTPN13 antibodies be used to study the protein's role in the mesenchymal-to-epithelial transition in cancer cells?

FITC-conjugated PTPN13 antibodies provide powerful tools for investigating the protein's role in mesenchymal-to-epithelial transition (MET) in cancer progression. Based on established research showing PTPN13's association with MET phenotype in xenograft models , the following methodological approach is recommended:

• Temporal expression analysis during MET:

  • Utilize time-course experiments with FITC-conjugated PTPN13 antibodies to track protein expression changes during induced MET

  • Implement live-cell imaging to monitor PTPN13 dynamics during junction formation in real-time

  • Correlate PTPN13 localization changes with cellular morphology transitions using phase contrast imaging

• Co-expression analysis with MET markers:

  • Design multi-color immunofluorescence panels combining FITC-conjugated PTPN13 antibodies with:

    • Epithelial markers: E-cadherin, cytokeratins, ZO-1

    • Mesenchymal markers: Vimentin, N-cadherin, Fibronectin

    • Transcription factors: SNAIL, SLUG, ZEB1, TWIST

  • Quantify co-expression patterns at single-cell resolution using high-content imaging systems

• Functional correlation studies:

  • Implement PTPN13 overexpression in aggressive mesenchymal lines (e.g., MDA-MB-231) and track:

    • Cell motility changes (wound healing assays)

    • Invasion capacity alterations (Boyden chamber assays)

    • Intercellular adhesion formation (cell aggregation assays)

  • Compare wild-type PTPN13 versus catalytically inactive mutants to isolate phosphatase-dependent effects

• Pathway integration analysis:

  • Investigate PTPN13's relationship with established MET pathways:

    • YAP signaling, which regulates PTPN13 through miR-30a

    • PI3K/Akt pathway components, particularly IRS-1 phosphorylation status

    • HER2 signaling interactions, based on transgenic mouse models

• In vivo visualization:

  • Apply FITC-conjugated PTPN13 antibodies in xenograft tumor sections to analyze:

    • Spatial distribution at tumor invasion fronts

    • Correlation with metastatic potential

    • Association with desmosome formation in vivo

• Quantitative analysis methods:

  • Implement image analysis algorithms to quantify:

    • PTPN13 expression levels during MET progression

    • Subcellular redistribution patterns

    • Co-localization coefficients with junction proteins

This comprehensive approach leverages fluorescent antibody technology to elucidate PTPN13's mechanistic contributions to the MET phenotype, with direct implications for understanding metastatic processes and potential therapeutic interventions.

How can I resolve weak or absent signal issues when using FITC-conjugated PTPN13 antibodies?

When encountering weak or absent signals with FITC-conjugated PTPN13 antibodies, implement this systematic troubleshooting approach:

• Sample preparation issues:

  • Fixation problems: Overfixation can mask epitopes; reduce fixation time or switch from paraformaldehyde to milder fixatives like methanol

  • Antigen retrieval optimization: For FFPE tissues, test both TE buffer (pH 9.0) and citrate buffer (pH 6.0) as recommended for PTPN13 antibodies

  • Permeabilization efficiency: Increase concentration or duration of detergent treatment (Triton X-100, saponin) to improve antibody access to intracellular PTPN13

• Antibody-specific factors:

  • Concentration adjustment: Increase antibody concentration beyond standard ranges (try 1:25 for IF applications when standard 1:50-1:500 dilutions fail)

  • Incubation optimization: Extend primary antibody incubation to overnight at 4°C to enhance binding

  • Antibody denaturation: Verify antibody storage conditions; improper storage can reduce activity even within the 12-month stability window

  • FITC photobleaching: Minimize exposure to light during all procedures and consider using antifade mounting media

• Target expression considerations:

  • Expression level verification: Confirm PTPN13 expression in your samples; expression varies widely across cell types

  • Epitope accessibility: Try antibodies targeting different PTPN13 domains (AA 1-161, AA 250-500, AA 801-900, AA 1965-2173)

  • Post-translational modifications: Consider that phosphorylation may mask epitopes; test phosphatase treatment

• Technical enhancements:

  • Signal amplification systems: Implement tyramide signal amplification to boost FITC signal

  • Alternative detection: Compare results with unconjugated primary followed by highly sensitive fluorescent secondary antibodies

  • Confocal settings optimization: Adjust laser power, detector gain, and pinhole settings for optimal FITC visualization

  • Spectral unmixing: Use spectral detectors to separate FITC signal from tissue autofluorescence

• Validation approaches:

  • Positive control inclusion: Process known PTPN13-positive samples (MCF-7 cells or HeLa cells ) in parallel

  • Alternative applications: Confirm PTPN13 presence using Western blot (1:1000-1:6000 dilution) before attempting IF

  • Antibody functionality test: Verify FITC conjugate fluorescence using dot blot or direct fluorometry

• Equipment considerations:

  • Filter sets: Ensure microscope filter sets match FITC spectral properties (excitation ~495nm, emission ~520nm)

  • Detector sensitivity: Use high-sensitivity cameras or photomultiplier tubes for low abundance targets

This comprehensive troubleshooting approach addresses the full range of potential issues affecting FITC-conjugated PTPN13 antibody performance.

What strategies can be employed to minimize background when using FITC-conjugated PTPN13 antibodies in tissues with high autofluorescence?

High background is a common challenge when using FITC-conjugated antibodies, particularly in tissues with natural autofluorescence. The following comprehensive strategies can minimize this issue when working with PTPN13 detection:

• Sample preparation optimization:

  • Autofluorescence quenching: Pretreat tissues with:

    • 0.1-1% sodium borohydride for 10 minutes (reduces aldehyde-induced fluorescence)

    • 0.1-0.3% Sudan Black B in 70% ethanol for 20 minutes (quenches lipofuscin)

    • 10-100mM CuSO₄ in 50mM ammonium acetate buffer (reduces general autofluorescence)

  • Fixation modification: Minimize fixation time or switch to methanol fixation which typically produces less autofluorescence than aldehyde-based fixatives

  • Fresh tissue processing: When possible, use fresh frozen tissues rather than FFPE samples to reduce processing-induced autofluorescence

• Antibody application refinements:

  • Blocking enhancement: Implement multi-step blocking:

    • Serum block: 5-10% serum from species unrelated to primary and secondary antibodies

    • Protein block: 1-3% BSA or casein to reduce non-specific binding

    • Fc receptor block: Using commercial Fc receptor blocking reagents

  • Antibody titration: Determine precise optimal concentration through serial dilution testing beyond the 1:50-1:500 range recommended for PTPN13 antibodies

  • Buffer optimization: Add 0.1-0.3% Triton X-100 and 0.05-0.1% Tween-20 to antibody diluent to reduce non-specific hydrophobic interactions

• Fluorescence differentiation techniques:

  • Spectral imaging: Employ spectral detectors to separate FITC signal (peak ~520nm) from autofluorescence

  • Alternative fluorophores: Consider switching to fluorophores outside the autofluorescence spectrum:

    • Alexa Fluor 647-conjugated antibodies (far-red spectrum)

    • Quantum dots with narrow emission spectra

  • Time-gated detection: Utilize time-resolved fluorescence microscopy to separate FITC signal from shorter-lived autofluorescence

• Imaging and analysis strategies:

  • Linear unmixing: Apply computational algorithms to separate FITC signal from autofluorescence spectra

  • Background subtraction: Acquire images from unstained adjacent sections for digital subtraction

  • Autofluorescence reference channels: Image pure autofluorescence in channels outside FITC excitation/emission

  • DAPI co-staining: Use DAPI nuclear counterstain to provide structural reference and aid in distinguishing true signal

• Validation controls:

  • Isotype controls: Use FITC-conjugated isotype-matched IgG at identical concentration to determine non-specific binding levels

  • Absorption controls: Pre-absorb antibody with recombinant PTPN13 fusion protein to confirm specificity

  • Negative controls: Include known PTPN13-negative tissues to establish baseline autofluorescence levels

These strategies can be employed individually or in combination depending on the specific tissue type and severity of autofluorescence interference when detecting PTPN13.

What are the best practices for dual or multi-color immunofluorescence experiments involving FITC-conjugated PTPN13 antibodies?

Successful multi-color immunofluorescence experiments involving FITC-conjugated PTPN13 antibodies require careful planning and execution. Based on the protein's demonstrated roles in cell junction stabilization and tumor suppression , the following best practices ensure optimal results:

• Experimental design considerations:

  • Fluorophore selection: Pair FITC (excitation: 495nm, emission: 520nm) with spectrally distinct fluorophores:

    • Combine with far-red dyes (Cy5, Alexa 647) for maximum separation

    • Avoid rhodamine/Texas Red which may show bleed-through with FITC

    • Consider DAPI (blue) as a nuclear counterstain for spatial reference

  • Target compatibility: For PTPN13 co-localization studies, prioritize antibodies against:

    • Desmosomal proteins (demonstrated interaction partners)

    • IRS-1 and phospho-IRS-1 (Tyr612) (established downstream targets)

    • Cell junction components to visualize PTPN13's role in junction stabilization

• Antibody selection and validation:

  • Host species diversity: Choose primary antibodies from different host species to avoid cross-reactivity

  • Mono vs. polyclonal consideration: For PTPN13, polyclonal antibodies show good reactivity with human samples , but ensure other antibodies in the panel are from different species

  • Pre-testing: Validate each antibody individually before combining in multiplexed experiments

  • Direct vs. indirect detection: Consider using directly conjugated antibodies for all targets to eliminate secondary antibody cross-reactivity

• Protocol optimization:

  • Sequential staining: For challenging combinations, implement sequential rather than simultaneous staining:

    • Apply, detect, and block the first primary antibody

    • Follow with subsequent antibodies with intervening blocking steps

  • Antibody concentration balancing: Adjust each antibody's concentration independently to achieve balanced signal intensity

  • Antigen retrieval coordination: If different antigens require different retrieval methods, test compatibility or implement sequential retrieval procedures

  • Fixation compromise: Select fixation method optimal for preserving PTPN13 epitopes while maintaining reactivity with co-staining targets

• Image acquisition strategies:

  • Sequential scanning: Capture each fluorophore channel separately to prevent cross-talk

  • Compensation settings: Apply digital compensation for any spectral overlap, particularly between FITC and yellow-orange fluorophores

  • Exposure optimization: Set exposure times to prevent FITC photobleaching while maintaining detection sensitivity

  • Z-stack acquisition: For colocalization analysis, collect optical sections at optimal z-resolution

• Controls and validation:

  • Single-color controls: Prepare samples with each antibody alone to establish bleed-through parameters

  • Fluorescence minus one (FMO) controls: Omit one primary antibody at a time to determine contribution to each channel

  • Colocalization quantification: Use appropriate statistical methods (Pearson's coefficient, Manders' overlap) to quantify PTPN13 colocalization with junction proteins

• Analysis considerations:

  • 3D reconstruction: For complex structures like cell junctions, implement 3D reconstruction from z-stacks

  • Colocalization mapping: Generate pixel-by-pixel colocalization maps between PTPN13 and junction proteins

  • Time-series analysis: For dynamic studies, implement drift correction and intensity normalization to account for photobleaching

These comprehensive guidelines ensure reliable multi-color imaging results when investigating PTPN13's functional interactions with other cellular components.

How is PTPN13 being investigated as a potential biomarker or therapeutic target in cancer research?

PTPN13's emerging role as both a biomarker and potential therapeutic target in cancer research reflects its complex functions in tumor suppression. Current research directions include:

These research directions highlight PTPN13's potential as both a clinically relevant biomarker and therapeutic target, with particular significance in understanding and potentially intervening in cancer invasion and metastasis mechanisms .

What recent technological advances have improved the detection and analysis of PTPN13 in complex biological samples?

Recent technological advances have significantly enhanced our ability to detect, visualize, and analyze PTPN13 in complex biological contexts. These innovations expand research capabilities beyond traditional applications of FITC-conjugated antibodies:

• Advanced microscopy techniques:

  • Super-resolution microscopy: Techniques such as STORM, PALM, and SIM now enable visualization of PTPN13 localization with 10-20nm resolution, revealing precise subcellular distribution at cell junctions that was previously undetectable with conventional fluorescence microscopy

  • Expansion microscopy: Physical expansion of specimens allows standard confocal microscopes to achieve effective super-resolution imaging of PTPN13 in relation to junction proteins

  • Light-sheet microscopy: Enables rapid, high-resolution 3D imaging of PTPN13 distribution in larger tissue volumes with minimal photobleaching of FITC conjugates

• Multiplexed detection systems:

  • Mass cytometry (CyTOF): Metal-tagged antibodies against PTPN13 and dozens of other proteins enable simultaneous quantification in single cells without fluorescence spectral limitations

  • Multiplexed ion beam imaging (MIBI): Allows detection of 40+ proteins including PTPN13 in tissue sections with subcellular resolution

  • Cyclic immunofluorescence (CycIF): Sequential staining and imaging cycles enable visualization of PTPN13 alongside 30-40 other proteins in the same sample

• Antibody engineering improvements:

  • Recombinant antibody fragments: Single-chain variable fragments (scFvs) and nanobodies against PTPN13 provide superior tissue penetration and reduced background

  • Site-specific conjugation: Precisely controlled FITC attachment preserves antibody affinity while maximizing fluorophore activity

  • Bivalent detection systems: Primary detection with unconjugated antibodies followed by fluorescent secondary Fab fragments improves signal-to-noise ratio

• Spatial -omics integration:

  • Spatial transcriptomics: Correlation of PTPN13 protein localization with spatially resolved mRNA expression

  • Digital spatial profiling: Quantitative analysis of PTPN13 alongside hundreds of proteins in spatially resolved regions of interest

  • In situ sequencing: Combined protein and RNA detection allows correlation of PTPN13 protein with its transcriptional context

• Artificial intelligence applications:

  • Automated image analysis: Deep learning algorithms for unbiased quantification of PTPN13 expression patterns across large datasets

  • Pattern recognition: Neural networks trained to recognize specific PTPN13 distribution patterns associated with cancer progression

  • Predictive modeling: Integration of PTPN13 expression data with clinical outcomes to develop predictive models

• Live-cell analysis innovations:

  • Fluorescent protein fusions: CRISPR knock-in of fluorescent tags enables real-time visualization of endogenous PTPN13 dynamics

  • Biosensors: FRET-based approaches to monitor PTPN13 phosphatase activity in living cells

  • Optogenetic control: Light-inducible PTPN13 activation systems to study temporal aspects of signaling

These technological advances collectively enable more precise, comprehensive analysis of PTPN13's dynamic behavior and functional interactions, particularly in the context of its tumor suppressor role and junction stabilization functions .

What are the emerging research questions about PTPN13's role in cellular signaling networks beyond cancer biology?

While PTPN13's tumor suppressor functions are well-documented , emerging research is exploring its broader roles in cellular signaling networks. These developing research questions represent important directions for FITC-conjugated PTPN13 antibody applications:

• Neurobiological functions:

  • What role does PTPN13 play in Schwann cell biology beyond proliferation regulation? Recent evidence indicates YAP regulation of Schwann cell proliferation and death is mediated through miR-30a regulation of PTPN13 , raising questions about:

    • PTPN13's potential involvement in myelination processes

    • Its role in Schwann cell responses to nerve injury

    • Possible contributions to neurodegenerative conditions

  • How does PTPN13 influence neuronal connectivity? Given its junction-stabilizing properties , researchers are investigating:

    • PTPN13's potential role in synapse formation and stability

    • Its contribution to neurodevelopmental processes

    • Possible dysregulation in neurological disorders

• Immunological significance:

  • What is PTPN13's role in immune cell function? Its original identification as APO-1/CD95 (Fas)-associated phosphatase suggests unexplored implications in:

    • T-cell activation and apoptosis regulation

    • Inflammatory signaling cascades

    • Autoimmune disease mechanisms

  • How does PTPN13 modulate the tumor immune microenvironment? Beyond direct effects on cancer cells , questions include:

    • Impact on tumor-infiltrating lymphocyte function

    • Influence on immunosuppressive cell recruitment

    • Potential modulation of immunotherapy responses

• Developmental biology questions:

  • What developmental processes require PTPN13 activity? Its role in cell adhesion suggests importance in:

    • Epithelial-mesenchymal transitions during development

    • Tissue morphogenesis and organogenesis

    • Stem cell niche establishment and maintenance

• Metabolic signaling interactions:

  • How does PTPN13's regulation of IRS-1 impact metabolic homeostasis? The established connection between PTPN13 and IRS-1 phosphorylation raises questions about:

    • Potential roles in insulin signaling beyond cancer

    • Contributions to metabolic syndrome pathophysiology

    • Interactions with nutrient-sensing pathways

• Cellular stress response regulation:

  • What is PTPN13's role in cellular adaptation to stress? Emerging questions include:

    • Involvement in mechanical stress sensing via cell junctions

    • Potential regulation of oxidative stress responses

    • Participation in ER stress and unfolded protein response pathways

• Intercellular communication mechanisms:

  • How does PTPN13 influence cell-cell communication beyond junction stabilization? Research is exploring:

    • Potential regulation of extracellular vesicle content and release

    • Influence on paracrine signaling pathways

    • Modulation of cell-cell communication in tissue microenvironments

These emerging research directions highlight PTPN13's potential significance in diverse biological contexts beyond its established cancer-related functions. FITC-conjugated PTPN13 antibodies provide valuable tools for investigating these questions through high-resolution visualization of protein localization and dynamics across different cellular contexts and model systems.

What are the key considerations when selecting the optimal PTPN13 antibody formulation for specific research applications?

Selecting the optimal PTPN13 antibody formulation requires careful consideration of multiple factors to ensure successful experimental outcomes. Based on comprehensive analysis of the available literature and technical specifications, researchers should consider:

• Application-specific requirements:

  • For live-cell applications: FITC-conjugated antibodies offer direct visualization without secondary detection steps, but may require higher concentrations than stated ranges for unconjugated antibodies (1:50-1:500)

  • For fixed-tissue work: Consider epitope sensitivity to fixation; both TE buffer (pH 9.0) and citrate buffer (pH 6.0) have proven effective for PTPN13 antigen retrieval

  • For multiplexing: Select FITC conjugates compatible with other fluorophores that minimize spectral overlap, especially when investigating PTPN13's co-localization with junction proteins

• Target epitope selection:

  • Different antibodies target distinct regions of the 2485 amino acid PTPN13 protein , including:

    • N-terminal regions (AA 1-161): Suitable for detecting full-length protein

    • Mid-region domains (AA 250-500, AA 801-900): May access different conformational states

    • C-terminal regions (AA 1965-2173): Include the catalytic phosphatase domain

  • Epitope accessibility varies by application; for instance, the phosphatase domain may be masked in protein complexes at cell junctions

• Species reactivity considerations:

  • Human reactivity: Most extensively validated in cancer research applications

  • Mouse reactivity: Important for transgenic models demonstrating PTPN13's tumor suppressor functions

  • Rat reactivity: Available for neurobiological research, particularly relevant for Schwann cell studies

  • Cross-reactivity testing: Always validate with appropriate positive controls for each species

• Technical performance parameters:

  • Signal-to-noise ratio: Polyclonal antibodies may offer higher sensitivity but potentially higher background

  • Specificity validation: Confirm target specificity through knockout/knockdown controls

  • Lot-to-lot consistency: Consider monoclonal options for long-term studies requiring consistent detection

  • Storage stability: Standard storage at -20°C provides 12-month stability; after reconstitution, aliquoting is recommended for conjugated formats

• Experimental design integration:

  • Quantitative applications: Select antibodies validated for flow cytometry with established titration protocols

  • Spatial analysis: Choose formulations validated for immunofluorescence with demonstrated subcellular localization patterns

  • Functional studies: Consider antibodies validated in conjunction with PTPN13 activity assays