PCED1A Antibody

What is PCED1A and why are antibodies against it important for research?

PCED1A (PC-esterase domain containing 1A) is a protein encoded by the PCED1A gene located on chromosome 20 in humans. While the complete function of PCED1A remains under investigation, research involving PCED1A antibodies has become increasingly important for studying its expression patterns in normal and pathological tissues, particularly in cancer research .

Antibodies against PCED1A are crucial research tools that allow for:

• Detection and quantification of PCED1A protein expression

• Examination of subcellular localization

• Investigation of protein-protein interactions

• Analysis of post-translational modifications

• Study of PCED1A's role in various cellular processes and disease states

The growing interest in PCED1A stems from observations of its differential expression in various cancer types, making antibodies against this protein valuable tools for oncology research .

What types of PCED1A antibodies are commercially available?

Based on current data, there are approximately 97 PCED1A antibodies available from 17 different providers . These antibodies can be categorized as follows:

When selecting an antibody, researchers should consider validation status, as several antibodies have published references supporting their specificity and utility .

What are the most commonly used applications for PCED1A antibodies?

PCED1A antibodies have been validated for multiple applications in research settings:

• Immunohistochemistry (IHC): For detecting PCED1A expression in tissue sections, particularly in cancer tissue analysis

• Immunocytochemistry (ICC): For examining subcellular localization in cultured cells

• Western Blotting (WB): For protein detection and semi-quantitative analysis of expression levels

• ELISA: For quantitative measurement of PCED1A protein levels in various sample types

• Immunofluorescence: Particularly with fluorophore-conjugated antibodies like FITC-labeled PCED1A antibodies for fluorescence microscopy applications

The choice of application should be guided by the specific research question and experimental design, with consideration of which antibodies have been validated for that particular application.

What are the optimal protocols for using PCED1A antibodies in Western blotting?

Based on available information, researchers should consider the following protocol guidelines for Western blotting using PCED1A antibodies:

• Sample preparation: Standard cell or tissue lysis with RIPA buffer containing protease inhibitors

• Protein loading: 20-40 μg of total protein per well is typically sufficient

• Antibody dilution: For polyclonal antibodies like those available from antibodies-online (ABIN405985), a recommended dilution of 1:1000 is typically effective

• Incubation conditions:

  • Primary antibody: Overnight at 4°C

  • Secondary antibody: 1 hour at room temperature

• Detection system: Both chemiluminescence and fluorescence-based detection systems are compatible

• Expected band size: Verify the expected molecular weight of PCED1A (~35-40 kDa depending on isoform and post-translational modifications)

For optimal results, researchers should include appropriate positive controls and validate antibody specificity using knockdown or knockout controls when possible.

How should PCED1A antibodies be validated for research applications?

Proper validation of PCED1A antibodies is critical for generating reliable research data. A comprehensive validation approach should include:

• Specificity testing:

  • Western blot analysis showing a single band at the expected molecular weight

  • Testing in multiple cell lines with known PCED1A expression levels

  • Knockdown/knockout validation (siRNA, CRISPR-Cas9) to confirm specificity

  • Peptide competition assays

• Application-specific validation:

  • For IHC: Testing on positive and negative control tissues

  • For ICC: Co-localization with known markers or GFP-tagged PCED1A

  • For ELISA: Standard curve generation and spike-recovery experiments

• Cross-reactivity assessment:

  • Testing reactivity across species (human, mouse, rat) when working with animal models

  • Evaluation of potential cross-reactivity with similar protein family members

• Lot-to-lot consistency evaluation:

  • Testing new lots against previously validated lots

  • Monitoring for changes in performance over time

Researchers should note that according to available data, several PCED1A antibodies have references supporting their validation, which can guide selection .

What are the optimal conditions for immunohistochemistry with PCED1A antibodies?

For optimal immunohistochemical detection of PCED1A in tissue samples, researchers should consider the following protocol guidelines:

• Fixation and processing:

  • Formalin-fixed paraffin-embedded (FFPE) sections (4-6 μm thick)

  • Antigen retrieval: Citrate buffer (pH 6.0) or EDTA buffer (pH 9.0), heat-mediated

• Blocking:

  • 5-10% normal serum (matched to secondary antibody host) with 1% BSA

  • 30-60 minutes at room temperature

• Primary antibody:

  • Recommended dilutions vary by product (1:100-1:500 for polyclonal antibodies like Invitrogen PA5-62248 or Novus Biologicals NBP2-13976)

  • Incubation: Overnight at 4°C or 1-2 hours at room temperature

• Detection systems:

  • Standard DAB (3,3'-diaminobenzidine) detection systems

  • Polymer-based detection systems for enhanced sensitivity

• Controls:

  • Positive control tissues: According to Human Protein Atlas data, include tissues with known PCED1A expression

  • Negative controls: Primary antibody omission and isotype controls

For multiplex IHC applications, researchers should validate antibody compatibility with multiplexing reagents and ensure minimal cross-reactivity with other antibodies in the panel.

How can PCED1A antibodies be used in cancer research?

PCED1A has shown differential expression patterns across various cancer types, making PCED1A antibodies valuable tools in oncology research . Key applications include:

• Expression analysis in tumor tissues:

  • Immunohistochemical staining of tumor microarrays to evaluate expression patterns across cancer types

  • Correlation of expression levels with clinical outcomes and survival data

  • Potential use as a prognostic or diagnostic biomarker

• Functional studies:

  • Investigation of protein interactions in cancer signaling pathways

  • Analysis of subcellular localization changes during cancer progression

  • Evaluation of post-translational modifications in tumor versus normal tissue

• Therapeutic target assessment:

  • Evaluation of PCED1A as a potential therapeutic target

  • Analysis of expression changes in response to treatment

  • Development of antibody-drug conjugates or targeted therapies

The Human Protein Atlas data indicates variable expression of PCED1A across different cancer types, suggesting tissue-specific roles that warrant further investigation .

What is known about PCED1A expression patterns in normal and disease tissues?

Based on available data from tissue expression studies:

• Normal tissue expression:

  • PCED1A shows variable expression across normal human tissues

  • Expression patterns can be assessed using validated antibodies for immunohistochemistry

  • The Human Protein Atlas provides a comprehensive analysis of PCED1A expression across normal tissues

• Cancer tissue expression:

  • Differential expression has been observed across various cancer types

  • Expression patterns may correlate with clinical outcomes in certain cancer types

  • Kaplan-Meier survival analysis has been performed for cancers showing significant PCED1A expression

• Other disease associations:

  • Limited data is available regarding PCED1A expression in non-cancer pathologies

  • Research is ongoing regarding potential roles in other disease states

Researchers should note that expression patterns should be validated across multiple antibodies when possible, especially when studying tissues not previously characterized for PCED1A expression.

How do PCED1A expression levels correlate with disease progression or prognosis?

The Human Protein Atlas data suggests potential prognostic significance of PCED1A expression in certain cancer types :

• Prognostic correlations:

  • High or low expression of PCED1A has shown significant (p<0.001) association with patient survival in specific cancer types

  • These associations may be either favorable or unfavorable depending on the cancer type

  • Kaplan-Meier survival analysis can be performed using appropriate tissue microarrays and validated antibodies

• Expression changes during disease progression:

  • Studies examining expression across different cancer stages or grades are ongoing

  • PCED1A antibodies can be used for temporal analysis of expression changes during disease progression

  • Correlation with other established biomarkers may provide insights into disease mechanisms

• Methodological considerations:

  • Quantitative analysis methods (H-score, digital image analysis) should be employed for reliable correlation studies

  • Multiple antibodies should be used to validate findings

  • Multivariate analysis should account for confounding clinical factors

Researchers should consider that prognostic significance may vary by cancer type, patient population, and treatment regimen.

How can PCED1A antibodies be used in conjunction with other research techniques?

PCED1A antibodies can be integrated with multiple advanced research techniques for comprehensive analysis:

• Multi-omics approaches:

  • Correlation of protein expression (immunohistochemistry) with transcriptomic data

  • Integration with proteomic profiling data

  • Correlation with genomic alterations or epigenetic modifications

• Advanced imaging techniques:

  • Super-resolution microscopy for detailed subcellular localization

  • Multiplex immunofluorescence for co-localization with other proteins

  • Live cell imaging with fluorescently-tagged antibody fragments

• Protein interaction studies:

  • Co-immunoprecipitation followed by mass spectrometry

  • Proximity ligation assays to detect protein-protein interactions in situ

  • ChIP-seq for potential transcription factor activity

• Functional studies:

  • Antibody-mediated inhibition experiments

  • Correlation with cellular phenotypes following genetic manipulation

  • Antibody-based protein purification for enzymatic or structural studies

These integrated approaches provide more comprehensive insights than single-method studies and help validate findings across multiple platforms.

What are the challenges in detecting post-translational modifications of PCED1A?

Detecting post-translational modifications (PTMs) of PCED1A presents several methodological challenges:

• Limited availability of PTM-specific antibodies:

  • Few validated antibodies exist that specifically recognize phosphorylated, glycosylated, or otherwise modified PCED1A

  • Development and validation of modification-specific antibodies require:

    • Synthetic modified peptides as immunogens

    • Extensive specificity testing against unmodified protein

    • Validation in biological systems with known modification states

• Technical considerations:

  • Sample preparation must preserve labile modifications

  • Enrichment strategies may be necessary for low-abundance modified forms

  • Appropriate controls (phosphatase treatment, deglycosylation, etc.) are essential

• Analytical approaches:

  • Mass spectrometry-based approaches may complement antibody-based detection

  • Combination of immunoprecipitation with modification-specific detection methods

  • Correlation with known modification-inducing conditions or treatments

For researchers interested in PCED1A modifications, developing a custom modification-specific antibody may be necessary, or alternatively, using mass spectrometry-based approaches following immunoprecipitation with total PCED1A antibodies.

How can PCED1A antibodies be used to study protein-protein interactions?

Investigating PCED1A protein interactions requires sophisticated approaches combining antibody-based techniques with other methods:

• Co-immunoprecipitation (Co-IP):

  • Using validated PCED1A antibodies to pull down protein complexes

  • Western blot or mass spectrometry analysis of co-precipitated proteins

  • Reciprocal Co-IP validation of identified interactions

  • Controls should include IgG control and ideally PCED1A-depleted samples

• Proximity ligation assay (PLA):

  • In situ detection of protein-protein interactions with <40 nm proximity

  • Requires validated antibodies against both PCED1A and putative interaction partners

  • Provides spatial information about interaction sites within cells

  • Quantitative analysis possible with appropriate image analysis tools

• Immunofluorescence co-localization:

  • Simultaneous detection of PCED1A and interaction partners

  • Super-resolution microscopy for detailed spatial analysis

  • Quantitative co-localization analysis using appropriate software

• Functional validation approaches:

  • Mutational analysis of interaction domains

  • Competition experiments with peptide fragments

  • Correlation with functional readouts

When designing interaction studies, researchers should consider cellular context, potential dynamic or transient interactions, and appropriate validation strategies.

What are common technical challenges when working with PCED1A antibodies and how can they be addressed?

Researchers may encounter several technical challenges when working with PCED1A antibodies:

• High background in immunostaining:

  • Solution: Optimize blocking conditions (try different blocking agents, increase blocking time)

  • Solution: Titrate antibody concentration

  • Solution: Increase washing stringency (more washes, higher salt concentration)

  • Solution: Use alternative detection systems with lower background

• Inconsistent or weak signal in Western blots:

  • Solution: Optimize protein extraction methods to preserve epitope integrity

  • Solution: Increase protein loading or antibody concentration

  • Solution: Extend primary antibody incubation time or use more sensitive detection systems

  • Solution: Verify protein transfer efficiency with reversible staining

• Cross-reactivity issues:

  • Solution: Validate specificity using knockout/knockdown controls

  • Solution: Perform peptide competition assays

  • Solution: Try alternative antibodies recognizing different epitopes

  • Solution: Increase washing stringency

• Batch-to-batch variability:

  • Solution: Test new lots against previously validated lots

  • Solution: Maintain consistent protocol parameters

  • Solution: Consider developing a standard operating procedure with positive controls

• Species cross-reactivity limitations:

  • Solution: Verify sequence homology at the epitope region across species

  • Solution: Test antibodies specifically validated for your species of interest

  • Solution: Consider custom antibody development for specific research needs

When troubleshooting, systematic modification of one variable at a time while maintaining appropriate controls is recommended for identifying optimal conditions.

How should researchers interpret conflicting results from different PCED1A antibodies?

When faced with discrepant results using different PCED1A antibodies, researchers should follow this systematic approach:

• Evaluate antibody validation status:

  • Review validation data for each antibody

  • Prioritize results from antibodies with more extensive validation

  • Consider the epitope location for each antibody (different domains may show different patterns)

• Perform additional validation experiments:

  • Test antibodies on known positive and negative controls

  • Validate with genetic approaches (siRNA, CRISPR knockout)

  • Perform peptide competition assays

  • Consider orthogonal detection methods (mass spectrometry)

• Consider biological explanations:

  • Different antibodies may recognize different isoforms

  • Post-translational modifications may mask epitopes

  • Protein conformation or interactions may affect epitope accessibility

  • Subcellular localization may affect detection

• Reporting recommendations:

  • Clearly document all antibodies used (catalog numbers, lots)

  • Report all conflicting results transparently

  • Discuss potential biological explanations for discrepancies

  • Consider using multiple antibodies and reporting consensus findings

When publishing research using PCED1A antibodies, researchers should provide complete methodological details and acknowledge potential limitations of antibody-based detection.

What quality control measures should researchers implement when working with PCED1A antibodies?

Implementing robust quality control measures is essential for generating reliable data with PCED1A antibodies:

• Antibody validation controls:

  • Positive and negative control samples with known PCED1A expression

  • Genetic validation (siRNA knockdown, CRISPR knockout)

  • Peptide competition assays

  • Orthogonal validation with alternative detection methods

• Experimental controls:

  • Technical replicates to assess reproducibility

  • Biological replicates to account for natural variation

  • Loading/normalization controls for quantitative analyses

  • Isotype controls for immunostaining

• Protocol standardization:

  • Documented standard operating procedures

  • Consistent lot usage when possible

  • Regular calibration of equipment

  • Consistent sample preparation methods

• Data analysis quality control:

  • Blinded quantification where appropriate

  • Standardized image acquisition parameters

  • Validated analysis software and algorithms

  • Statistical validation of findings

• Documentation practices:

  • Detailed record-keeping of reagent information

  • Comprehensive methodological documentation

  • Raw data preservation

  • Transparent reporting of limitations

Implementing these quality control measures increases data reliability and reproducibility, which is particularly important when working with antibody-based detection methods.

What emerging applications for PCED1A antibodies show promise for future research?

Several emerging applications for PCED1A antibodies show potential for advancing understanding of this protein:

• Single-cell analysis:

  • Integration with single-cell technologies to analyze expression heterogeneity

  • Spatial transcriptomics combined with protein detection

  • Single-cell Western blotting for quantitative analysis at cellular level

• Therapeutic applications:

  • Development of function-blocking antibodies if PCED1A shows therapeutic relevance

  • Antibody-drug conjugates for targeted therapy if overexpressed in specific cancers

  • Companion diagnostics for patient stratification

• Structural biology applications:

  • Antibody-assisted cryo-EM for structural determination

  • Epitope mapping to identify functional domains

  • Conformation-specific antibodies to detect structural changes

• In vivo imaging:

  • Development of imaging probes based on validated antibodies

  • Non-invasive detection of PCED1A expression in animal models

  • Correlation with disease progression in preclinical models

• Systems biology approaches:

  • Integration with multi-omics datasets

  • Network analysis of protein interactions

  • Correlation with cellular phenotypes and disease states

Researchers interested in these emerging applications should consider collaborative approaches with specialists in these respective fields.

How can researchers contribute to improving PCED1A antibody validation and standardization?

Researchers can significantly contribute to improving PCED1A antibody validation through several approaches:

• Comprehensive validation studies:

  • Systematic testing across multiple applications

  • Cross-comparison of different commercial antibodies

  • Publication of detailed validation data

  • Development of standard validation protocols

• Data sharing initiatives:

  • Contribution to antibody validation repositories

  • Sharing of positive and negative control samples

  • Publication of detailed protocols and methodologies

  • Reporting of both successful and unsuccessful applications

• Development of reference standards:

  • Creation of recombinant PCED1A standards

  • Development of cell/tissue reference materials

  • Establishment of quantitative benchmarks

  • Generation of knockout/knockdown validation materials

• Collaborative research networks:

  • Multi-laboratory validation studies

  • Inter-laboratory protocol standardization

  • Shared resource development

  • Consensus guideline development

• Technical innovations:

  • Development of improved antibody formats (recombinant, fragments)

  • Creation of modification-specific antibodies

  • Integration with emerging technologies

  • Improvement of detection sensitivity and specificity

By contributing to these efforts, researchers can help advance the reliability and utility of PCED1A antibodies for the broader scientific community.