XPNPEP1 Antibody

What is XPNPEP1 and what cellular functions does it perform?

XPNPEP1, also known as X-prolyl aminopeptidase 1 or soluble aminopeptidase P (sAmp), is a cytosolic metalloaminopeptidase that catalyzes the cleavage of N-terminal amino acids adjacent to proline residues. The enzyme plays significant roles in the degradation and maturation of tachykinins, neuropeptides, and peptide hormones . As a soluble form of aminopeptidase P, XPNPEP1 is primarily expressed in the cytosol, with notable expression in pancreatic and intestinal tissues . This 70-75 kDa protein (observed molecular weight) is encoded by the XPNPEP1 gene, which can undergo alternative splicing to produce multiple transcript variants . Understanding this protein's function is crucial for researchers studying proteolytic pathways and neuropeptide processing.

What types of XPNPEP1 antibodies are available for research use?

Several types of XPNPEP1 antibodies are available for research applications:

These antibodies vary in their specificity, sensitivity, and applications, allowing researchers to select the most appropriate tool for their experimental needs . When selecting an antibody, consider the target species, application requirements, and whether a monoclonal or polyclonal antibody would better suit your research objectives.

What are the recommended applications for XPNPEP1 antibodies?

XPNPEP1 antibodies have been validated for multiple research applications with specific recommended dilutions:

It is recommended to titrate the antibody concentration for optimal results in each specific experimental system . Positive tissue controls for WB include mouse/rat pancreas and small intestine tissues, while for IHC, human pancreas and small intestine tissues have shown positive results .

How should I optimize Western blot protocols for XPNPEP1 detection?

For optimal Western blot detection of XPNPEP1:

• Sample preparation: Use fresh tissue samples from pancreas or small intestine for highest yield. Lysates should be prepared with protease inhibitors to prevent degradation.

• Gel selection: Use 10-12% polyacrylamide gels to properly resolve the 70-75 kDa XPNPEP1 protein.

• Transfer conditions: Optimize transfer time based on protein size; typically 60-90 minutes at 100V for proteins in this molecular weight range.

• Antibody dilution: Begin with 1:2000 dilution of primary antibody (e.g., 10661-1-AP) and titrate as needed for optimal signal-to-noise ratio .

• Incubation conditions: For primary antibody, incubate overnight at 4°C with gentle agitation to improve specificity and reduce background.

• Detection system: HRP-conjugated secondary antibodies with ECL detection systems work well for XPNPEP1 visualization.

• Positive controls: Include mouse pancreas tissue or mouse small intestine tissue as positive controls to validate antibody performance .

Remember that the observed molecular weight (70-75 kDa) may vary slightly from the calculated weight (70 kDa) due to post-translational modifications .

What are the critical factors for successful immunohistochemical detection of XPNPEP1?

Successful immunohistochemical detection of XPNPEP1 requires attention to several critical factors:

• Tissue fixation: Formalin-fixed, paraffin-embedded (FFPE) tissues are commonly used. Overfixation can mask epitopes, so standardize fixation time.

• Antigen retrieval: For XPNPEP1, TE buffer at pH 9.0 is the preferred antigen retrieval method, though citrate buffer at pH 6.0 can be used as an alternative .

• Blocking: Thorough blocking with appropriate serum (typically 5-10% normal serum from the same species as the secondary antibody) is essential to reduce non-specific binding.

• Antibody dilution: Start with 1:100 dilution for IHC and optimize based on staining intensity and background. The recommended range is 1:50-1:500 .

• Incubation conditions: Primary antibody incubation at 4°C overnight generally yields better results than shorter incubations at room temperature.

• Detection systems: DAB (3,3'-diaminobenzidine) chromogen provides good contrast for visualization. Amplification systems may be needed for low-abundance targets.

• Controls: Include known positive tissues (human pancreas or small intestine) and negative controls (primary antibody omission or isotype controls) .

• Counterstaining: Light hematoxylin counterstaining helps visualize tissue architecture without obscuring specific staining.

Validation through multiple antibodies or complementary techniques enhances confidence in IHC results.

How do I select the appropriate XPNPEP1 antibody for my specific research application?

Selecting the appropriate XPNPEP1 antibody involves considering several factors:

• Species reactivity: Confirm that the antibody recognizes XPNPEP1 in your experimental species. For example, 10661-1-AP reacts with human, mouse, and rat XPNPEP1, while some antibodies are human-specific .

• Application compatibility: Verify validation data for your intended application:

  • For Western blot: Antibodies like 10661-1-AP have been validated at 1:1000-1:4000 dilutions

  • For IHC: Several antibodies work well, with recommended dilutions between 1:50-1:500

  • For ELISA or other specialized applications: Check specific validation data

• Monoclonal vs. polyclonal:

  • Monoclonal antibodies (like MA5-47892) offer higher specificity for a single epitope

  • Polyclonal antibodies (like 10661-1-AP) recognize multiple epitopes, potentially increasing sensitivity but with higher background risk

• Epitope location: Consider whether your research requires detection of a specific region or isoform of XPNPEP1.

• Validation data: Review published validation data, including:

  • Western blot bands at expected molecular weight (70-75 kDa)

  • IHC staining patterns in appropriate tissues

  • ELISA binding characteristics

• Research question: Match antibody characteristics to your specific research questions. For example, if studying protein-protein interactions, an antibody that doesn't interfere with interaction domains would be preferable.

Creating a comparison table of available antibodies with their specific characteristics can facilitate this selection process.

What are common issues when using XPNPEP1 antibodies and how can they be resolved?

When troubleshooting, it's advisable to run multiple controls, including positive tissue controls (pancreas and small intestine samples) and negative controls (irrelevant antibodies or tissues not expressing XPNPEP1) .

How can I validate the specificity of my XPNPEP1 antibody?

Validating the specificity of XPNPEP1 antibodies should involve multiple complementary approaches:

• Western blot validation:

  • Confirm single band at expected molecular weight (70-75 kDa)

  • Test multiple tissues known to express XPNPEP1 (pancreas, small intestine)

  • Include negative controls (tissues with low/no expression)

  • Consider knockdown/knockout validation if available

• Peptide competition assay:

  • Pre-incubate antibody with immunizing peptide

  • Specific signal should be significantly reduced or eliminated

  • Non-specific binding will remain

• Multiple antibody comparison:

  • Test different antibodies targeting distinct epitopes of XPNPEP1

  • Consistent results across antibodies increase confidence in specificity

  • Compare polyclonal (10661-1-AP) and monoclonal (MA5-47892) antibodies

• Immunoprecipitation followed by mass spectrometry:

  • Confirm pulled-down protein is indeed XPNPEP1

  • Identify any co-precipitating proteins that might cause cross-reactivity

• Correlation with mRNA expression:

  • Compare protein detection with RT-PCR or RNA-seq data

  • Expression patterns should correlate across tissues

• Functional validation:

  • For neutralizing antibodies, demonstrate inhibition of enzymatic activity

  • For detection antibodies, correlate signal with known biology

Documenting these validation steps is essential for publishing reliable research results using XPNPEP1 antibodies .

What are the optimal storage and handling conditions for XPNPEP1 antibodies?

To maintain antibody performance and extend shelf life, observe these storage and handling recommendations:

• Temperature conditions:

  • Store antibodies at -20°C for long-term storage

  • Avoid repeated freeze-thaw cycles by preparing working aliquots

  • For antibodies like 10661-1-AP, they remain stable for one year when properly stored

• Buffer composition:

  • Most XPNPEP1 antibodies are supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3

  • This formulation helps maintain stability during freeze-thaw cycles

  • Some preparations may contain BSA (e.g., 0.1%) as a stabilizer

• Aliquoting strategy:

  • Upon receipt, prepare small working aliquots (10-20 μL)

  • For 20 μL sizes containing 0.1% BSA, aliquoting is unnecessary for -20°C storage

  • Label aliquots with antibody details, date, and dilution information

• Working solution handling:

  • Dilute antibodies immediately before use

  • Keep diluted antibodies on ice when in use

  • Discard diluted antibodies after 1-2 days or as recommended

• Contamination prevention:

  • Use sterile pipette tips and tubes

  • Avoid introducing bacteria or fungi

  • Never return unused antibody to the original stock

• Transport considerations:

  • Transport on ice or with cold packs

  • Minimize time at room temperature

  • Check for signs of degradation upon arrival (precipitation, color change)

Proper documentation of antibody lot numbers, storage conditions, and thawing cycles helps track performance and troubleshoot inconsistencies between experiments.

How can XPNPEP1 antibodies be utilized in multiplex immunoassays or Immuno-MRM approaches?

XPNPEP1 antibodies can be effectively incorporated into advanced multiplexed detection systems:

• Immuno-MRM (Mass Spectrometry-based Immunoassay):

  • The CPTC-XPNPEP1-1 antibody has been validated for Immuno-MRM applications, particularly in plasma samples

  • This technique combines antibody-based enrichment with mass spectrometry detection

  • Workflow involves:
    a) Antibody capture of XPNPEP1 from complex samples
    b) Enzymatic digestion of captured proteins
    c) MS detection of specific peptides

  • Provides higher specificity than traditional immunoassays by verifying protein identity through peptide mass

  • Enables absolute quantification when combined with isotope-labeled standards

• Multiplex immunoassays:

  • XPNPEP1 can be incorporated into bead-based multiplex systems, such as the NexaTag™ qIPCR ELISA Kit

  • These systems allow simultaneous detection of multiple proteins from a single sample

  • Key considerations include:
    a) Antibody cross-reactivity with other targets in the panel
    b) Dynamic range compatibility between targets
    c) Buffer conditions suitable for all included antibodies

• Chromatin immunoprecipitation (ChIP) adaptations:

  • Modified protocols can use XPNPEP1 antibodies to study protein-DNA interactions

  • Particularly relevant when investigating XPNPEP1's potential role in transcriptional regulation

• Proximity ligation assays (PLA):

  • Can detect protein-protein interactions involving XPNPEP1

  • Requires careful selection of antibody pairs targeting different epitopes

  • Provides spatial information about protein interactions in situ

• Single-cell proteomics applications:

  • Emerging technologies allow XPNPEP1 detection at single-cell resolution

  • Requires highly specific antibodies with minimal background

When designing multiplex assays, researchers must validate that the XPNPEP1 antibody maintains specificity and sensitivity in the multiplex environment, as other components may affect binding characteristics .

What are the considerations for using XPNPEP1 antibodies in studying protein-protein interactions?

When studying XPNPEP1 protein-protein interactions, researchers should consider:

• Antibody epitope location:

  • Ensure the antibody binding site doesn't interfere with interaction domains

  • Map the epitope recognized by the antibody to avoid potential steric hindrance

  • For immunoprecipitation applications, verify the antibody doesn't compete with interaction partners

• Co-immunoprecipitation (Co-IP) optimization:

  • Buffer conditions must preserve native protein structure and interactions

  • Mild detergents (0.1% NP-40 or Triton X-100) help maintain interactions

  • Salt concentration should be optimized (typically 100-150 mM NaCl)

  • Pre-clearing lysates reduces non-specific binding

• Cross-linking considerations:

  • Reversible cross-linkers can stabilize transient interactions

  • Cross-linking conditions must be optimized to avoid artificial aggregation

  • Carefully control cross-linking time and concentration

• Reciprocal confirmation:

  • Verify interactions by immunoprecipitating with antibodies against interaction partners

  • Results should be consistent regardless of which protein is used as the bait

• Controls for specificity:

  • Include IgG controls from the same species as the antibody

  • Include samples where XPNPEP1 is downregulated or absent

  • Competitive peptide controls can confirm antibody specificity

• Compatible detection methods:

  • Consider using antibodies from different species for immunoprecipitation vs. detection

  • Alternatively, use conjugated primary antibodies to avoid interference from IP antibodies

  • Mass spectrometry can identify novel interaction partners without specific antibodies

• Validation through complementary methods:

  • Confirm interactions using multiple techniques (Co-IP, PLA, FRET)

  • Functional assays should demonstrate biological relevance of identified interactions

Understanding XPNPEP1's role in degrading bioactive peptides makes protein-protein interaction studies particularly relevant for identifying substrates and regulatory partners.

How can XPNPEP1 antibodies be employed in quantitative proteomic studies?

XPNPEP1 antibodies can be integrated into several quantitative proteomic workflows:

• Immuno-MRM (Multiple Reaction Monitoring):

  • CPTC-XPNPEP1-1 antibody has been successfully used in Immuno-MRM applications

  • This technique enables absolute quantification of XPNPEP1 in complex samples

  • Workflow involves:
    a) Immunoaffinity enrichment of XPNPEP1
    b) Tryptic digestion to generate target peptides
    c) LC-MS/MS analysis with MRM to quantify specific peptides

  • Benefits include high specificity, wide dynamic range, and multiplexing capability

• Reverse Phase Protein Arrays (RPPA):

  • XPNPEP1 antibodies can be applied to high-throughput quantification across many samples

  • Requires highly specific antibodies with minimal cross-reactivity

  • Sample preparation must ensure consistent protein extraction

• Quantitative immunohistochemistry:

  • Digital image analysis of IHC staining can provide semi-quantitative data

  • Standardization with calibration controls is essential

  • Considers factors like staining intensity and percentage of positive cells

• ELISA-based quantification:

  • NexaTag™ qIPCR ELISA Kit for XPNPEP1 combines ELISA specificity with PCR sensitivity

  • Provides quantitative measurements in serum, plasma, and cell culture supernatants

  • Standard curves allow determination of absolute concentration

• Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA):

  • Combines isotope-labeled peptide standards with anti-peptide antibodies

  • Enables precise absolute quantification

  • Requires development of specific anti-peptide antibodies

• Considerations for experimental design:

  • Establish linear range of detection for selected method

  • Include appropriate calibration standards

  • Account for matrix effects in complex biological samples

  • Validate results with orthogonal quantification methods

• Data analysis approaches:

  • Normalization strategies to account for technical variation

  • Statistical methods appropriate for the experimental design

  • Integration with other -omics data for systems biology approaches

For accurate quantification, researchers must thoroughly validate antibody specificity and develop robust standard curves using recombinant XPNPEP1 protein or calibrated reference materials .

What are the latest developments in using XPNPEP1 antibodies for biomarker discovery and validation?

Recent advances in XPNPEP1 antibody applications for biomarker research include:

• Circulating XPNPEP1 detection:

  • Development of highly sensitive immunoassays capable of detecting XPNPEP1 in plasma and serum samples

  • The NexaTag™ Human Aminopeptidase P1 qIPCR ELISA Kit merges sandwich ELISA specificity with PCR sensitivity for improved detection limits

  • Potential applications in monitoring XPNPEP1 levels as biomarkers for specific pathological conditions

• Tissue-specific expression profiling:

  • Antibody-based characterization of XPNPEP1 expression across normal and diseased tissues

  • IHC validation through the Human Protein Atlas using antibodies like CPTC-XPNPEP1-1

  • Correlation of expression patterns with clinical outcomes to assess biomarker utility

• Post-translational modification analysis:

  • Development of antibodies specific to modified forms of XPNPEP1

  • Investigation of how PTMs affect XPNPEP1 function and potentially serve as more specific biomarkers

  • Correlation of modification status with disease progression or treatment response

• Multiplex biomarker panels:

  • Integration of XPNPEP1 detection into multiplex panels with other biomarkers

  • Immuno-MRM approaches using validated antibodies like CPTC-XPNPEP1-1 for multiplexed detection

  • Improved diagnostic accuracy through combinatorial biomarker signatures

• Extracellular vesicle analysis:

  • Examining XPNPEP1 content in exosomes and other extracellular vesicles

  • Potential for liquid biopsy applications using antibody-based capture and detection

• Digital pathology integration:

  • Combining XPNPEP1 IHC with digital image analysis for quantitative assessment

  • Machine learning algorithms to identify subtle expression patterns associated with disease

• Validation methodologies:

  • Multi-institutional validation studies to establish reproducibility

  • Rigorous analytical validation following FDA and CLIA guidelines for potential clinical applications

  • Standardization of pre-analytical variables affecting XPNPEP1 measurement

These developments highlight the evolving role of XPNPEP1 antibodies in translational research, moving from basic characterization to potential clinical applications in diagnostics and personalized medicine .

What is the recommended protocol for using XPNPEP1 antibodies in Western blotting?

Detailed Western Blot Protocol for XPNPEP1 Detection:

• Sample preparation:

  • Harvest cells or tissues (pancreas and small intestine show strong expression)

  • Lyse in RIPA buffer containing protease inhibitors

  • Sonicate briefly to shear DNA and reduce viscosity

  • Centrifuge at 14,000 × g for 15 minutes at 4°C

  • Determine protein concentration (BCA or Bradford assay)

• SDS-PAGE:

  • Prepare 10% SDS-PAGE gel (optimal for 70-75 kDa XPNPEP1)

  • Load 20-50 μg protein per lane

  • Include molecular weight marker and positive control (pancreas tissue lysate)

  • Run at 100V until dye front reaches bottom of gel

• Transfer:

  • Transfer to PVDF membrane at 100V for 90 minutes or 30V overnight at 4°C

  • Verify transfer efficiency with Ponceau S staining

• Blocking:

  • Block membrane in 5% non-fat dry milk in TBST for 1 hour at room temperature

• Primary antibody incubation:

  • Dilute XPNPEP1 antibody 1:2000 in 5% BSA in TBST

  • Incubate overnight at 4°C with gentle agitation

• Washing:

  • Wash 3 × 10 minutes with TBST

• Secondary antibody incubation:

  • Incubate with HRP-conjugated anti-rabbit IgG (1:5000) in 5% non-fat dry milk in TBST for 1 hour at room temperature

• Final washing and detection:

  • Wash 3 × 10 minutes with TBST

  • Apply ECL substrate and detect signal

  • Expected band: 70-75 kDa

• Validation controls:

  • Positive control: Mouse or rat pancreas tissue lysate

  • Negative control: Tissue known to express minimal XPNPEP1

  • Loading control: Probe for housekeeping protein (e.g., GAPDH, β-actin)

For troubleshooting, common issues include weak signal (increase protein loading or antibody concentration), high background (increase blocking or reduce antibody concentration), or multiple bands (optimize lysate preparation to prevent degradation) .

What is the detailed protocol for immunohistochemical detection of XPNPEP1 in tissue samples?

Comprehensive IHC Protocol for XPNPEP1 Detection:

• Tissue preparation:

  • Fix tissues in 10% neutral buffered formalin for 24-48 hours

  • Process and embed in paraffin

  • Section at 4-5 μm thickness onto positively charged slides

  • Dry sections overnight at 37°C

• Deparaffinization and rehydration:

  • Xylene: 3 × 5 minutes

  • 100% ethanol: 2 × 3 minutes

  • 95% ethanol: 1 × 3 minutes

  • 70% ethanol: 1 × 3 minutes

  • Distilled water: 1 × 5 minutes

• Antigen retrieval (critical step for XPNPEP1):

  • Primary method: TE buffer pH 9.0 (10 mM Tris, 1 mM EDTA)

  • Alternative method: Citrate buffer pH 6.0

  • Heat in pressure cooker/microwave until boiling, then 15-20 minutes at sub-boiling temperature

  • Cool slides to room temperature (approximately 20 minutes)

  • Wash in PBS: 3 × 5 minutes

• Endogenous peroxidase blocking:

  • Incubate sections in 3% H₂O₂ in methanol for 10 minutes

  • Wash in PBS: 3 × 5 minutes

• Protein blocking:

  • Apply 5-10% normal goat serum in PBS for 30-60 minutes at room temperature

  • Drain blocking solution (do not wash)

• Primary antibody incubation:

  • Dilute XPNPEP1 antibody in antibody diluent (1:100 is recommended starting dilution, range: 1:50-1:500)

  • Incubate overnight at 4°C in humidified chamber

  • Wash in PBS: 3 × 5 minutes

• Detection system:

  • Apply HRP-polymer conjugated secondary antibody for 30 minutes at room temperature

  • Wash in PBS: 3 × 5 minutes

  • Develop with DAB chromogen for 5-10 minutes (monitor microscopically)

  • Wash in distilled water

• Counterstaining and mounting:

  • Counterstain with Harris hematoxylin for 30 seconds

  • Rinse in running tap water

  • Blue in 0.2% ammonia water or lithium carbonate

  • Dehydrate through graded alcohols and xylene

  • Mount with permanent mounting medium

• Controls:

  • Positive tissue control: Human pancreas or small intestine tissue

  • Negative control: Primary antibody replaced with isotype-matched IgG

  • Internal control: Non-target tissues within the section

Expected results include cytoplasmic staining in relevant cell types, with pancreatic and intestinal tissues showing prominent XPNPEP1 expression . Document staining patterns with digital images, noting intensity, subcellular localization, and percentage of positive cells.

How can I develop a quantitative ELISA assay for XPNPEP1 using available antibodies?

Development of a Quantitative ELISA for XPNPEP1:

• Antibody selection strategy:

  • Choose capture antibody: Monoclonal antibody (e.g., CPTC-XPNPEP1-1) for high specificity

  • Choose detection antibody: Polyclonal antibody (e.g., 10661-1-AP) that recognizes different epitopes

  • Alternatively, use a commercial kit like the NexaTag™ Human Aminopeptidase P1 qIPCR ELISA for established protocols

• Basic sandwich ELISA protocol:

  • Plate coating:

    • Dilute capture antibody in coating buffer (50 mM carbonate-bicarbonate, pH 9.6)

    • Apply 100 μL per well to high-binding 96-well plate

    • Incubate overnight at 4°C

  • Blocking:

    • Wash 3× with PBST (PBS + 0.05% Tween-20)

    • Block with 300 μL 1-5% BSA in PBS for 1-2 hours at room temperature

  • Sample preparation:

    • Prepare standards from recombinant XPNPEP1 (serial dilutions)

    • Dilute samples appropriately in sample diluent

  • Sample incubation:

    • Add 100 μL standards and samples to appropriate wells

    • Incubate 2 hours at room temperature or overnight at 4°C

    • Wash 4-5× with PBST

  • Detection antibody:

    • Add 100 μL diluted detection antibody

    • Incubate 1-2 hours at room temperature

    • Wash 4-5× with PBST

  • Secondary antibody/detection:

    • Add 100 μL HRP-conjugated secondary antibody

    • Incubate 1 hour at room temperature

    • Wash 5× with PBST

    • Add 100 μL TMB substrate

    • Incubate 5-30 minutes protected from light

    • Stop reaction with 100 μL 2N H₂SO₄

    • Read absorbance at 450 nm (reference 570 nm)

• Assay optimization considerations:

  • Determine optimal antibody pair through checkerboard titration

  • Optimize sample diluent composition to minimize matrix effects

  • Establish standard curve range covering physiological concentrations (typically pg/mL to ng/mL)

  • Determine limits of detection and quantification

  • Assess intra- and inter-assay variation (aim for CV < 15%)

• Validation steps:

  • Specificity: Confirm using recombinant XPNPEP1, knockout samples, or immunodepletion

  • Linearity: Serial dilutions of samples should yield proportional results

  • Recovery: Spike-in experiments with known quantities of recombinant protein

  • Precision: Multiple measurements of the same samples

  • Stability: Assess sample stability under various storage conditions

• Enhanced sensitivity options:

  • Amplification systems (e.g., biotin-streptavidin)

  • Chemiluminescent substrates instead of colorimetric

  • Advanced detection methods like the qIPCR approach used in NexaTag™ kits

By following these guidelines, researchers can develop a reliable quantitative ELISA for XPNPEP1 measurement across various sample types, including serum, plasma, and cell culture supernatants .

What are key publications that have utilized XPNPEP1 antibodies in significant research findings?

While specific publications are not directly cited in the search results, researchers investigating XPNPEP1 should be aware of the foundational research in this field and methodological papers demonstrating antibody applications. Key areas of XPNPEP1 research include its role in:

• Peptide hormone metabolism: Studies examining XPNPEP1's function in cleaving N-terminal amino acids adjacent to proline residues in bioactive peptides

• Enzymatic characterization: Research characterizing XPNPEP1's metalloaminopeptidase activity and substrate specificity

• Tissue expression profiling: Comprehensive studies documenting XPNPEP1 expression across tissues, with notable expression in pancreas and intestinal tissues

• Alternative splicing: Investigations of multiple transcript variants resulting from alternative splicing of the XPNPEP1 gene

• Methodological developments: Papers describing Immuno-MRM approaches for XPNPEP1 detection, as validated with antibodies like CPTC-XPNPEP1-1

The Human Protein Atlas has incorporated XPNPEP1 antibodies in their systematic protein expression mapping efforts, providing valuable reference data for tissue expression patterns . Additionally, the CPTAC (Clinical Proteomic Tumor Analysis Consortium) has utilized antibodies like CPTC-XPNPEP1-1 in cancer proteomics studies .

Researchers should conduct literature searches in PubMed and other scientific databases using terms like "XPNPEP1," "aminopeptidase P," or "X-prolyl aminopeptidase" to identify the most recent and relevant publications for their specific research questions.

Where can researchers access additional validation data for commercial XPNPEP1 antibodies?

Researchers seeking comprehensive validation data for XPNPEP1 antibodies can access information from several sources:

• Manufacturer validation galleries:

  • Proteintech provides validation data for antibody 10661-1-AP, including Western blot and IHC results from multiple tissues

  • Thermo Fisher Scientific offers validation information for their XPNPEP1 antibodies (MA5-47892 and PA5-19139)

• Antibody validation repositories:

  • Human Protein Atlas: Contains validation data for XPNPEP1 antibodies, including CPTC-XPNPEP1-1, with tissue expression profiles and IHC images

  • Antibodypedia: Aggregates validation data from multiple sources

• CPTAC Assay Portal (assays.cancer.gov):

  • Contains detailed validation data for CPTC-XPNPEP1-1 (CPTAC-694) with Immuno-MRM results

  • Includes plasma validation data, demonstrating utility in clinical samples

• Research Resource Identifiers (RRID) portal:

  • Using RRIDs such as AB_2215792 for 10661-1-AP or AB_2617378 for CPTC-XPNPEP1-1

  • Provides centralized information and citations for specific antibodies

• Publications and protocols:

  • Product-specific protocols are available for applications like Western blot and IHC

  • Published literature using specific XPNPEP1 antibodies often contains detailed methodological validation

• Collaborative research groups:

  • The CPTAC antibody portal (antibodies.cancer.gov) provides additional information on antibodies used in cancer proteomics research

  • Fred Hutchinson Cancer Research Center (Paulovich Lab) provides Immuno-MRM validation data

When evaluating validation data, researchers should consider:

• Application-specific validation (WB, IHC, ELISA)

• Species cross-reactivity testing

• Positive and negative controls used

• Quantitative metrics (sensitivity, specificity, dynamic range)

• Batch-to-batch consistency information

Accessing comprehensive validation data helps researchers select the most appropriate antibody for their specific experimental needs and anticipate potential limitations.

How might new antibody technologies enhance XPNPEP1 research in the coming years?

Emerging antibody technologies are poised to transform XPNPEP1 research in several ways:

• Next-generation recombinant antibodies:

  • Development of synthetic antibodies with enhanced specificity and reproducibility

  • CRISPR-engineered antibody-producing cell lines for consistent manufacturing

  • Standardized production methods to eliminate batch-to-batch variation observed in traditional antibodies

• Single-domain antibodies and nanobodies:

  • Smaller antibody fragments that can access epitopes unavailable to conventional antibodies

  • Enhanced penetration into tissues and subcellular compartments

  • Potential for improved detection of XPNPEP1 in its native cellular context

• Spatially resolved proteomics:

  • Integration of XPNPEP1 antibodies into multiplex tissue imaging platforms

  • Technologies like Imaging Mass Cytometry, CODEX, or GeoMx DSP to visualize XPNPEP1 in spatial context

  • Correlation of XPNPEP1 distribution with other proteins in tissue microenvironments

• Engineered affinity reagents:

  • Aptamer-based alternatives to traditional antibodies

  • Molecularly imprinted polymers (MIPs) designed to recognize specific XPNPEP1 epitopes

  • Synthetic binding proteins with tailored affinity and specificity

• Dynamic proteomics:

  • Time-resolved antibody-based assays to track XPNPEP1 dynamics

  • Photoswitchable antibodies or binding probes for super-resolution microscopy

  • Live-cell compatible nanobodies for tracking XPNPEP1 in real-time

• Antibody-drug conjugates (ADCs):

  • If XPNPEP1 shows disease-specific expression patterns, ADCs could enable targeted therapeutic approaches

  • Research tools combining antibody specificity with functional payloads to manipulate XPNPEP1 activity

• AI-enhanced antibody design:

  • Machine learning algorithms predicting optimal epitopes for XPNPEP1 targeting

  • In silico antibody engineering to maximize specificity and minimize cross-reactivity

  • Digital twins of antibody-antigen interactions to predict performance

These technological advances will likely enhance the specificity, sensitivity, and reproducibility of XPNPEP1 detection, enabling more sophisticated functional studies and potentially revealing new biological roles for this aminopeptidase.

What are potential future applications of XPNPEP1 antibodies in clinical diagnostics or therapeutics?

While current XPNPEP1 antibodies are primarily for research use only, their potential translation to clinical applications presents several promising avenues:

• Diagnostic biomarker development:

  • Integration into multiplex protein panels for disease detection

  • Quantitative assays like the NexaTag™ Human Aminopeptidase P1 qIPCR ELISA Kit could be adapted for clinical diagnostics

  • Liquid biopsy applications measuring XPNPEP1 in circulation as a disease indicator

• Companion diagnostics:

  • If XPNPEP1 activity is associated with drug metabolism or efficacy

  • Patient stratification based on XPNPEP1 expression or activity levels

  • Monitoring treatment response through changes in XPNPEP1 expression

• Histopathological applications:

  • Incorporation into diagnostic IHC panels for tissue classification

  • Digital pathology algorithms integrating XPNPEP1 staining patterns

  • Automated scoring systems for standardized interpretation

• Therapeutic antibody development:

  • Neutralizing antibodies targeting XPNPEP1 enzymatic activity

  • Antibody-drug conjugates if XPNPEP1 shows disease-specific upregulation

  • Bispecific antibodies linking XPNPEP1 to immune effector cells

• Theranostic approaches:

  • Dual-purpose antibodies for simultaneous imaging and therapy

  • XPNPEP1-targeted nanoparticles for drug delivery and imaging

  • Radio-immunoconjugates for targeted radiotherapy

• Point-of-care testing:

  • Adaptation of XPNPEP1 immunoassays to lateral flow or microfluidic platforms

  • Rapid testing applications if XPNPEP1 emerges as an acute biomarker

  • Integration with smartphone-based readers for decentralized diagnostics

• Circulating enzyme activity monitoring:

  • Development of activity-based probes conjugated to XPNPEP1 antibodies

  • Real-time monitoring of enzymatic function in patient samples

  • Correlation of enzyme activity with disease progression or therapeutic response