DPEP1 Antibody, Biotin conjugated

What is DPEP1 and why is it significant in biomedical research?

Dipeptidase 1 (DPEP1) is a zinc-dependent metalloproteinase that plays multiple crucial roles in cellular physiology. It belongs to the peptidase M19 family and functions in the hydrolysis of various dipeptides, particularly in renal metabolism. DPEP1 is significant because:

• It processes various peptides and antibiotics including thienamycin, penem, and carbapenem derivatives

• It participates in glutathione and leukotriene metabolism, converting leukotriene D4 to leukotriene E4

• It serves as a regulator of inflammatory mediators including those involved in asthma, arthritis, and inflammatory bowel disease

• It has been identified as a marker for colorectal cancer progression

• Recent research has discovered it functions as a receptor for coronavirus PHEV

The protein has a molecular weight of approximately 46 kDa and is anchored to cell membranes, making it accessible for antibody-based detection methods in various tissues.

What are the key specifications of commercially available DPEP1 Antibody, Biotin conjugated?

The DPEP1 Antibody, Biotin conjugated reagents typically have the following specifications:

These specifications are critical for determining compatibility with your experimental system and achieving reproducible results .

How should DPEP1 Antibody, Biotin conjugated be stored and handled to maintain optimal activity?

Proper storage and handling of the DPEP1 Antibody, Biotin conjugated is essential for maintaining its binding specificity and signal strength:

• Store at -20°C or -80°C immediately upon receipt

• Aliquot the antibody into smaller volumes before freezing to avoid repeated freeze/thaw cycles, which can significantly degrade antibody performance

• When creating aliquots, use sterile technique and sterile tubes

• Before use, allow the antibody to equilibrate to room temperature gradually

• Avoid vortexing the antibody as this can cause denaturation; instead, mix by gentle inversion or flicking

• The buffer contains 50% glycerol which helps prevent freezing damage, but proper aliquoting is still recommended

• Working dilutions should be prepared fresh before each experiment

• The antibody maintains stability for approximately one year when stored properly

Methodologically, it's advisable to run controls with each experiment to verify antibody performance, as even properly stored antibodies can lose activity over time.

What experimental techniques are compatible with DPEP1 Antibody, Biotin conjugated?

While DPEP1 Antibody, Biotin conjugated is primarily validated for ELISA applications, researchers should consider the following methodological approaches for various experimental contexts:

For each application, appropriate positive and negative controls should be included. For colorectal cancer studies, HCT-15 cells have been identified as DPEP1-positive, while specific knock-out or knock-down cell lines would serve as ideal negative controls .

How can I optimize DPEP1 Antibody, Biotin conjugated for immunohistochemistry in cancer tissue samples?

Optimizing DPEP1 Antibody, Biotin conjugated for IHC in cancer tissues requires careful consideration of several methodological factors:

• Tissue preparation and fixation:

  • FFPE (formalin-fixed paraffin-embedded) tissues have been successfully used for DPEP1 detection

  • A notable methodological finding is that DPEP1 detection was achieved without prior antigen retrieval, which differs from many other antibodies

• Staining protocol optimization:

  • Begin with a 1:500 dilution as reported in literature for DPEP1 antibodies

  • Use a biotin-based detection system (e.g., avidin-biotin-peroxidase complex)

  • Block endogenous biotin in tissues (particularly important in liver, kidney, and breast samples)

  • Consider using tyramide signal amplification for enhanced sensitivity

• Scoring system development:

  • Implement a three-tier system (weak, moderate, or strong) for evaluating DPEP1 expression

  • Count cells as positive when at least 10% show DPEP1 staining

  • For tissue microarrays, take the modal score when staining differs between cores

  • When the modal score is not applicable, the highest score should be used

• Validation approaches:

  • Include known positive tissues (colorectal cancer samples) and negative controls

  • Consider double staining with proliferation markers like Ki67 to correlate DPEP1 expression with cell cycle activity

This methodological approach has allowed researchers to identify DPEP1 as a marker for the transition from low-grade to high-grade intraepithelial neoplasia in colorectal cancer progression .

What are the recommended protocols for using DPEP1 Antibody, Biotin conjugated in ELISA assays?

For optimal performance in ELISA applications, consider the following methodological guidelines:

Direct ELISA Protocol:

• Coat microtiter plate wells with sample containing DPEP1 (typically 1-10 μg/ml in carbonate buffer, pH 9.6)

• Incubate overnight at 4°C

• Block with 1-5% BSA in PBST for 1-2 hours at room temperature

• Add biotin-conjugated DPEP1 antibody at empirically determined dilution

• Incubate for 1-2 hours at room temperature

• Wash 3-5 times with PBST

• Add streptavidin-HRP (typically 1:1000-1:5000 dilution)

• Incubate for 30-60 minutes at room temperature

• Wash 3-5 times with PBST

• Add TMB substrate and monitor color development

• Stop reaction with 2N H₂SO₄ or 1N HCl

• Read absorbance at 450nm

Sandwich ELISA Considerations:

• For sandwich ELISA, use a capture antibody against a different epitope of DPEP1

• The biotin-conjugated DPEP1 antibody serves as the detection antibody

• Detection ranges of 0.156-10 ng/mL have been reported for DPEP1 ELISA systems

Competitive ELISA Application:

• Pre-incubate samples with biotin-conjugated DPEP1 antibody

• Add this mixture to wells coated with recombinant DPEP1

• Higher concentrations of DPEP1 in samples will result in lower signal

• Standard curves should be generated using purified recombinant DPEP1 (23-252 AA)

For all ELISA formats, optimization of antibody concentration through titration is essential for maximizing signal-to-noise ratio and ensuring assay sensitivity.

How can DPEP1 Antibody, Biotin conjugated be utilized in cancer biomarker research?

Research has established DPEP1 as a significant biomarker in cancer progression, particularly in colorectal cancer. Methodological approaches for utilizing the biotin-conjugated antibody in this context include:

• Prognostic stratification studies:

  • Implement IHC analysis of tissue microarrays with standardized scoring systems

  • Correlate DPEP1 expression with histological staging parameters

  • Construct receiver operating characteristic (ROC) curves to evaluate prognostic value

  • Research has demonstrated DPEP1 as an independent adverse prognostic factor with ROC-AUC values of 0.9230 in colorectal cancer

• Early neoplastic lesion detection:

  • Focus on the transition from low-grade to high-grade intraepithelial neoplasia (IEN)

  • Develop graduated scoring systems for detecting subtle changes in DPEP1 expression

  • Combine with other markers to create biomarker panels with enhanced specificity

  • Research methodology has confirmed striking upregulation of DPEP1 in high-grade compared to low-grade IEN

• Multiplex biomarker systems:

  • Combine DPEP1 antibody detection with other cancer markers

  • Implement double immunostaining with proliferation markers (e.g., Ki67)

  • Use multiplexed immunofluorescence with streptavidin-conjugated fluorophores

  • Quantify co-localization using digital image analysis platforms

• Detection in liquid biopsies:

  • Develop protocols for DPEP1 detection in blood, serum, or plasma samples

  • Utilize the biotin-conjugated antibody in ELISA systems optimized for biological fluids

  • Combine with exosome isolation for enhanced sensitivity

  • Detection ranges of 0.156-10 ng/mL provide suitable sensitivity for many biological samples

These methodological approaches leverage the high specificity of DPEP1 antibodies to advance cancer biomarker research beyond simple detection to clinically relevant prognostic and diagnostic applications.

What role does DPEP1 play in coronavirus research, and how can the antibody be utilized in viral studies?

Recent groundbreaking research has identified DPEP1 as a functional receptor for porcine hemagglutinating encephalomyelitis virus (PHEV), opening new avenues for coronavirus research. Methodological approaches for utilizing DPEP1 Antibody, Biotin conjugated in this context include:

• Viral receptor binding studies:

  • Use the antibody to block DPEP1-virus interactions

  • Implement competition assays between soluble DPEP1 and the antibody

  • Develop flow cytometry protocols to quantify binding of viral particles to cell surface DPEP1

  • Research has shown that soluble DPEP1 strongly inhibits PHEV pseudotype infection with IC50 values of 0.03-0.04 μg/mL

• Receptor expression profiling in susceptible tissues:

  • Map DPEP1 expression patterns across tissues using the biotin-conjugated antibody

  • Correlate expression levels with susceptibility to viral infection

  • Implement double-staining with viral proteins to visualize co-localization

  • Both human and porcine DPEP1 orthologs allow viral entry, suggesting conservation of this mechanism

• Spike protein interaction mechanism studies:

  • Develop ELISA-based binding assays using the biotin-conjugated antibody

  • Implement binding inhibition studies to map interaction domains

  • Analyze whether the antibody interferes with spike-mediated cell-cell fusion

  • Research has determined that the enzymatic activity of DPEP1 is dispensable for viral entry, suggesting a structural role in receptor function

• Antiviral therapeutic development:

  • Screen for compounds that disrupt DPEP1-virus interactions

  • Use the antibody as a positive control for inhibition studies

  • Develop quantitative assays for receptor occupancy

  • Studies show that preincubation with soluble DPEP1 reduced virus release by 2.3-3.5 fold in infected cells

This methodological approach represents a cutting-edge application of DPEP1 antibodies in understanding novel virus-host interactions, with potential implications for coronavirus pathogenesis beyond PHEV.

How can potential cross-reactivity and specificity issues with DPEP1 Antibody, Biotin conjugated be addressed in complex experimental systems?

Addressing specificity concerns requires rigorous validation strategies:

• Comprehensive validation approach:

  • Implement knockout/knockdown controls using CRISPR-Cas9 or siRNA

  • Compare reactivity patterns with multiple antibodies against different DPEP1 epitopes

  • Verify molecular weight (46 kDa) in immunoblotting applications

  • Consider preabsorption tests with recombinant DPEP1 protein (23-252 AA)

• Species cross-reactivity considerations:

  • While primarily validated against human DPEP1, testing in other species requires validation

  • Sequence homology analysis between human DPEP1 and orthologs should guide experimental design

  • The porcine ortholog shares functional conservation with human DPEP1 in virus binding studies

  • Document any differences in signal intensity or localization patterns between species

• Isoform-specific detection:

  • Consider potential reactivity with DPEP1 isoforms or related family members

  • The immunogen (23-252 AA) should be evaluated for sequence uniqueness

  • UniProt database lists secondary accession numbers D3DX80 and Q96AK2, which may represent variant forms

  • Design experiments to differentiate between closely related peptidases

• Tissue-specific interference mitigation:

  • Address potential interference from endogenous biotin in tissues like liver and kidney

  • Implement blocking steps using streptavidin/avidin before antibody application

  • Consider alternative detection systems for tissues with high background

  • Test multiple sample preparation methods to maximize signal-to-noise ratio

• Quantitative validation metrics:

  • Establish signal-to-noise ratios for different applications

  • Document antibody titration curves to determine optimal concentration

  • Implement appropriate statistical analyses to distinguish specific from non-specific signals

  • Consider multiplexed detection to correlate DPEP1 signal with known co-markers

This comprehensive approach ensures that experimental findings attributed to DPEP1 detection are truly specific and not the result of antibody cross-reactivity with related proteins or non-specific binding.

What are common challenges when using DPEP1 Antibody, Biotin conjugated, and how can they be addressed?

Researchers frequently encounter several methodological challenges when working with DPEP1 Antibody, Biotin conjugated. Here are evidence-based solutions:

Systematic optimization through pilot experiments addressing each variable independently will yield the most reliable protocol for your specific research application.

How can researchers optimize detection sensitivity when working with samples containing low DPEP1 expression?

Optimizing detection of low-abundance DPEP1 requires methodological refinements at multiple experimental stages:

• Sample preparation enhancement:

  • Implement enrichment strategies for membrane proteins

  • Consider cell surface protein biotinylation followed by streptavidin pull-down

  • For tissue samples, use laser capture microdissection to isolate DPEP1-expressing cells

  • Optimize lysis buffers to effectively solubilize GPI-anchored proteins like DPEP1

• Signal amplification technologies:

  • Implement tyramide signal amplification (TSA) systems for immunohistochemistry

  • Utilize polymeric detection systems instead of traditional ABC methods

  • For ELISA applications, consider using poly-HRP conjugated streptavidin

  • Implement bioorthogonal amplification strategies such as rolling circle amplification

• Detection system optimization:

  • Extend primary antibody incubation times (overnight at 4°C)

  • Utilize high-sensitivity substrate systems (SuperSignal, ECL Prime)

  • Implement chemiluminescent substrates with extended signal duration

  • Consider digital image accumulation for weak signals

• Instrument sensitivity maximization:

  • For microplate readers, utilize extended integration times

  • Implement PMT gain optimization for flow cytometry applications

  • For microscopy, use high-sensitivity cameras with cooling systems

  • Consider spectral unmixing to separate signal from autofluorescence

• Validated sensitivity benchmarks:

  • DPEP1 ELISA systems have demonstrated sensitivity down to 0.094 ng/mL

  • Detection ranges of 0.156-10 ng/mL have been established for serum and plasma samples

  • For tissue expression analysis, implement standardized scoring systems with clear thresholds

By systematically implementing these methodological refinements, researchers can achieve detection of DPEP1 even in samples with low expression levels, enabling studies in contexts where DPEP1 dysregulation might be subtle but biologically significant.

What advanced methodological approaches can be used to study DPEP1 protein-protein interactions using the biotin-conjugated antibody?

Investigating DPEP1 protein-protein interactions requires sophisticated methodological approaches that leverage the biotin conjugation of the antibody:

• Proximity-based interaction studies:

  • Implement proximity ligation assays (PLA) to visualize interactions with <40nm resolution

  • Combine the biotin-conjugated DPEP1 antibody with antibodies against suspected interaction partners

  • Utilize PLA to investigate DPEP1 interactions with leukotrienes and carbapenem derivatives

  • Quantify interaction signals using appropriate image analysis software

• Co-immunoprecipitation strategies:

  • Use streptavidin-coated magnetic beads to pull down the biotin-conjugated antibody-DPEP1 complex

  • Analyze co-precipitated proteins by mass spectrometry

  • Implement stringent washing conditions to minimize non-specific interactions

  • Validate findings with reverse co-immunoprecipitation using antibodies against interaction partners

• Functional interaction mapping:

  • Implement competitive binding assays between DPEP1 and viral proteins like coronavirus spike

  • Develop dose-response curves for inhibition studies using recombinant proteins

  • Calculate binding parameters such as IC50 values (0.03-0.04 μg/mL reported for PHEV spike)

  • Use surface plasmon resonance to determine kinetic parameters of interactions

• In situ visualization techniques:

  • Perform dual-labeling immunofluorescence with DPEP1 and interaction partners

  • Implement FRET-based assays to confirm direct protein-protein interactions

  • Utilize super-resolution microscopy (STORM, PALM) to visualize nanoscale interaction domains

  • Quantify co-localization using appropriate statistical methods (Pearson's correlation, Manders' coefficients)

• Dynamic interaction studies:

  • Develop live-cell imaging protocols using the biotin-conjugated antibody and fluorescent streptavidin

  • Implement FRAP (Fluorescence Recovery After Photobleaching) to assess interaction dynamics

  • Utilize pulse-chase experiments to track temporal aspects of interactions

  • Consider microfluidic approaches for real-time monitoring of binding events

These advanced methodological approaches enable researchers to move beyond simple detection of DPEP1 to understanding its functional interactions in complex biological systems, particularly in contexts like viral binding and cancer progression where protein-protein interactions are mechanistically significant.

How is DPEP1 Antibody, Biotin conjugated being used in cutting-edge inflammation and leukocyte recruitment research?

Recent advancements have positioned DPEP1 as a significant target in inflammation research, with the biotin-conjugated antibody playing crucial methodological roles:

• Neutrophil recruitment mechanisms:

  • DPEP1 has been identified as an adhesion receptor for neutrophil recruitment in lungs and liver

  • The biotin-conjugated antibody enables precise localization of DPEP1 at inflammatory sites

  • Flow cytometric protocols using streptavidin-fluorophore detection systems quantify DPEP1 expression on neutrophils

  • Blocking experiments with the antibody help elucidate the mechanistic role of DPEP1 in neutrophil extravasation

• Anti-inflammatory therapeutic development:

  • Pharmaceutical compositions targeting DPEP1 are being developed to reduce inflammation

  • The antibody serves as a validation tool for compound screening assays

  • Competition assays between potential therapeutics and the antibody help identify binding sites

  • Patent applications describe DPEP1-binding compositions with potential for inflammatory disease treatment

• Leukotriene metabolism regulation:

  • DPEP1 converts leukotriene D4 to leukotriene E4, regulating inflammatory signaling

  • The biotin-conjugated antibody enables correlation of DPEP1 expression with leukotriene levels

  • Enzymatic activity assays combined with immunolocalization reveal functional domains

  • Inhibitor screening assays identify compounds that modulate DPEP1's metabolic functions

• Sepsis and acute kidney injury models:

  • DPEP1-targeting compositions show promise in treating sepsis-related inflammation

  • The antibody helps track DPEP1 distribution in kidney tissues during acute injury

  • Quantitative immunoassays monitor DPEP1 levels as biomarkers of renal inflammatory damage

  • Therapeutic approaches aim to block DPEP1-mediated leukocyte recruitment in acute scenarios

These cutting-edge applications demonstrate the evolving significance of DPEP1 beyond its classical roles in renal metabolism, positioning the biotin-conjugated antibody as an essential tool in inflammation research.

How can DPEP1 Antibody, Biotin conjugated be integrated into multi-omics research approaches?

Integration of DPEP1 antibody detection into multi-omics frameworks requires sophisticated methodological considerations:

• Proteogenomic integration strategies:

  • Correlate DPEP1 protein expression (detected via the antibody) with mRNA expression profiles

  • Implement CITE-seq approaches combining antibody detection with single-cell transcriptomics

  • Map DPEP1 expression to genomic variants that may affect protein abundance

  • Research has demonstrated significant correlation between DPEP1 protein expression and mRNA levels in colorectal cancer, with a ROC-AUC of 0.9230

• Glycoproteomics applications:

  • DPEP1 is a GPI-anchored glycoprotein identified in N-glycoproteome studies

  • The biotin-conjugated antibody can help verify accessibility of glycosylated epitopes

  • Combine with lectin-based enrichment to study DPEP1 glycoforms

  • N-glycosylation site identification dictates accessible epitopes for antibody binding

• Spatial multi-omics implementation:

  • Utilize the biotin-conjugated antibody in spatial proteomics platforms

  • Correlate DPEP1 localization with tissue metabolomics data

  • Implement digital spatial profiling to quantify DPEP1 in the context of the tissue microenvironment

  • Integrate with mass spectrometry imaging for comprehensive spatial mapping

• Systems biology framework development:

  • Position DPEP1 within protein interaction networks using antibody-based interaction studies

  • Correlate DPEP1 expression with metabolic profiles, particularly leukotriene and glutathione pathways

  • Develop mathematical models incorporating DPEP1 expression data from antibody-based assays

  • Implement network pharmacology approaches to predict impacts of DPEP1 modulation

This integrated approach elevates DPEP1 research from isolated protein studies to comprehensive biological understanding, positioning the biotin-conjugated antibody as a bridge between proteomics and other omics domains.

What are the emerging applications of DPEP1 Antibody, Biotin conjugated in developmental biology and stem cell research?

The involvement of DPEP1 in fundamental biological processes offers new avenues for developmental and stem cell research applications:

• Pluripotent stem cell surface marker studies:

  • DPEP1 has been identified in cell surface N-glycoproteome studies of human pluripotent stem cells

  • The biotin-conjugated antibody enables live cell sorting of stem cell populations

  • Flow cytometric protocols can identify developmental transitions based on DPEP1 expression

  • Methodologies for both positive and negative selection using DPEP1 as a marker have been validated

• Lineage differentiation tracking:

  • Monitor DPEP1 expression changes during differentiation of stem cells toward specific lineages

  • Develop quantitative immunofluorescence protocols for developmental time-course studies

  • Correlate DPEP1 expression with differentiation stage-specific markers

  • Implement sorting strategies to isolate DPEP1-expressing cells at critical developmental junctures

• Organoid development applications:

  • Apply the antibody in 3D culture systems to track DPEP1 expression in developing organoids

  • Establish protocols for whole-mount immunostaining of organoids with the biotin-conjugated antibody

  • Correlate DPEP1 expression patterns with functional maturation of kidney and intestinal organoids

  • Implement clearing techniques for deep tissue imaging of DPEP1 in complex 3D structures

• Reproductive and developmental toxicology:

  • Utilize DPEP1 detection as a marker for developmental toxicity in kidney and intestinal tissues

  • Develop high-throughput screening assays based on DPEP1 expression in embryonic tissues

  • Implement quantitative image analysis workflows for toxicity assessment

  • Correlate DPEP1 modulation with developmental outcomes in model organisms

These emerging applications highlight the utility of DPEP1 Antibody, Biotin conjugated beyond traditional research contexts, offering new methodological approaches for understanding developmental processes and stem cell biology.

What are the most promising future research directions for DPEP1 Antibody, Biotin conjugated applications?

Based on current research trends and emerging findings, several high-potential research directions stand out:

• Viral receptor inhibition studies:

  • The discovery of DPEP1 as a functional receptor for coronavirus PHEV opens unprecedented research avenues

  • Future applications will likely focus on developing therapeutic approaches using antibodies or small molecules

  • Comparative studies of DPEP1 binding across coronavirus species may reveal evolutionary adaptations

  • Methodological approaches combining structural biology with antibody-based functional studies will be crucial

• Cancer progression and metastasis mechanisms:

  • DPEP1's role in colorectal cancer progression suggests applications in other cancer types

  • The transition from biomarker to therapeutic target represents a significant opportunity

  • Combined approaches targeting both diagnosis and treatment based on DPEP1 expression

  • Development of immunotherapeutic approaches leveraging DPEP1 as a target on cancer cells

• Inflammatory disease intervention:

  • DPEP1's role in neutrophil recruitment and leukotriene metabolism positions it as a therapeutic target

  • Development of more specific inhibitors based on structural understanding of antibody binding sites

  • Refinement of biomarker applications in inflammatory conditions beyond current applications

  • Integration of DPEP1-targeting approaches with existing anti-inflammatory strategies

• Advanced imaging and detection technologies:

  • Development of multiplexed detection systems incorporating DPEP1 with other biomarkers

  • Implementation of emerging super-resolution techniques for nanoscale localization

  • Integration with mass cytometry for high-dimensional analysis of DPEP1 in complex tissues

  • Development of in vivo imaging approaches using modified antibody fragments

These diverse research directions highlight the versatility of DPEP1 Antibody, Biotin conjugated as a tool in biomedical research, with applications spanning from basic molecular understanding to clinical translation.

How might technological advances impact the utility and applications of DPEP1 antibodies in research?

Emerging technologies will transform how researchers utilize DPEP1 antibodies in several methodological dimensions:

• Single-cell and spatial biology integration:

  • Implementation of DPEP1 antibodies in single-cell proteomics platforms

  • Development of spatial proteomics approaches combining transcriptomics with DPEP1 protein detection

  • Integration with emerging spatial multi-omics platforms for comprehensive tissue analysis

  • Application in microfluidic single-cell analysis systems for high-throughput functional studies

• Artificial intelligence and computational biology:

  • Development of AI-assisted image analysis for quantifying DPEP1 expression in complex tissues

  • Implementation of machine learning algorithms for predicting DPEP1 interactions based on antibody binding data

  • Integration of structural biology predictions with antibody epitope mapping

  • Development of systems biology models incorporating DPEP1 functional data

• Antibody engineering advancements:

  • Creation of recombinant antibody formats with enhanced specificity and reduced background

  • Development of bi-specific antibodies targeting DPEP1 and interaction partners simultaneously

  • Implementation of antibody fragments for improved tissue penetration

  • Creation of switchable antibody systems for temporal control of DPEP1 detection

• Translational medicine applications:

  • Development of companion diagnostic applications based on DPEP1 detection

  • Implementation in liquid biopsy platforms for non-invasive cancer monitoring

  • Integration with point-of-care testing systems for inflammatory conditions

  • Development of theranostic approaches combining DPEP1 detection with therapeutic delivery