EDIL3 Antibody

What is EDIL3 and why is it important in research?

EDIL3, also known as DEL1 (Developmental endothelial locus-1), is an extracellular matrix protein secreted primarily by endothelial cells. This protein contains three epidermal growth factor (EGF) domains and two discoidin domains, playing critical roles in various physiological and pathological processes including embryonic development, angiogenesis, inflammation, and immune reactions . EDIL3 has gained significant research interest due to its involvement in cancer progression, where its expression is elevated in multiple cancer types including gastric cancer, hepatocellular carcinoma, pancreatic adenocarcinoma, colorectal cancer, and breast cancer . Its ability to promote endothelial cell adhesion through interactions with integrins, particularly alpha-v/beta-3 integrin receptor, makes it an important target for vascular biology and oncology research . The protein also functions as an endogenous inhibitor of inflammatory cell recruitment, further expanding its research significance in immunology .

How do I select the appropriate EDIL3 antibody for my experiment?

When selecting an EDIL3 antibody, consider these key experimental parameters:

• Application compatibility: Determine which application you need (Western blot, IHC, ELISA, etc.) and select an antibody validated for that application. For instance, ABIN2784767 is validated for Western blotting, while E-AB-64606 is validated for immunohistochemistry .

• Species reactivity: Match the antibody to your experimental model. Some antibodies like ABIN2784767 offer broad species reactivity (human, mouse, rat, cow, dog, guinea pig, rabbit, zebrafish, horse), while others may be more species-restricted .

• Epitope recognition: Consider which domain or region of EDIL3 you need to target:

  • N-terminal targeting antibodies like ABIN2784767

  • Specific amino acid region antibodies, such as those targeting AA 101-199, AA 337-365, or AA 60-300

• Clonality: Determine whether a polyclonal or monoclonal antibody better suits your needs:

  • Polyclonal antibodies (like E-AB-64606) offer broader epitope recognition

  • Monoclonal antibodies provide higher specificity for a single epitope

• Validation data: Review the validation methods used for the antibody, including predicted reactivity percentages and cross-reactivity testing .

What are the most common applications for EDIL3 antibodies?

EDIL3 antibodies are utilized across multiple experimental applications, each providing distinct insights into this protein's expression and function:

Each application requires specific optimization parameters including antibody dilution, incubation conditions, and signal detection methods .

How should I optimize Western blot protocols for EDIL3 detection?

EDIL3 detection via Western blotting requires careful optimization to ensure specific and sensitive results:

• Sample preparation:

  • Use appropriate lysis buffers containing protease inhibitors to prevent EDIL3 degradation

  • For secreted EDIL3, consider concentrating cell culture supernatants

• Protein loading:

  • Load 20-50 μg of total protein from cell/tissue lysates

  • For secreted EDIL3, concentration methods may be necessary

• Gel selection:

  • Use 8-10% SDS-PAGE gels for optimal resolution of the ~52 kDa EDIL3 protein

  • Consider gradient gels (4-15%) for simultaneous detection of different molecular weight proteins

• Antibody dilution:

  • Primary antibody: Start with manufacturer's recommended dilution (typically 1:1000 for ABIN2784767)

  • Secondary antibody: Use 1:5000-1:10000 dilution of HRP-conjugated secondary antibody

• Incubation conditions:

  • Primary antibody: Overnight at 4°C for optimal signal-to-noise ratio

  • Secondary antibody: 1 hour at room temperature

• Detection considerations:

  • EDIL3 may show different bands due to glycosylation or processing

  • Expected molecular weight is approximately 52 kDa, but may vary depending on post-translational modifications

  • Include positive controls (tissues known to express EDIL3, such as embryonic endothelial tissues)

• Troubleshooting:

  • If background is high, increase blocking time or try different blocking reagents

  • If no signal is detected, verify EDIL3 expression in your samples or use a positive control

What protocol adaptations are needed for EDIL3 immunohistochemistry?

For effective immunohistochemical detection of EDIL3 in tissue samples:

• Tissue preparation:

  • Use 10% neutral buffered formalin fixation (10-24 hours)

  • Paraffin embedding with standard protocols

  • Section tissues at 4-6 μm thickness

• Antigen retrieval:

  • Heat-induced epitope retrieval using citrate buffer (pH 6.0) is typically effective

  • Pressure cooking for 3-5 minutes or microwave heating for 10-20 minutes

• Antibody dilution:

  • For E-AB-64606: Use 1:50-1:200 dilution as recommended

  • For ab198003: Start with 1:20 dilution as validated for human cervical and ovarian cancer tissues

• Detection systems:

  • Use polymer-based HRP detection systems for improved sensitivity

  • DAB (3,3'-diaminobenzidine) is commonly used as chromogen

  • Hematoxylin counterstaining for nuclear visualization

• Controls:

  • Positive control: Human stomach tissue has been verified for E-AB-64606

  • Negative controls: Omit primary antibody or use isotype control

• Interpretation considerations:

  • EDIL3 is primarily a secreted protein, so expect extracellular matrix staining

  • Evaluate both staining intensity and pattern (membranous, cytoplasmic, or extracellular)

  • Consider quantitative scoring methods for comparative studies

How can I validate EDIL3 antibody specificity?

Ensuring antibody specificity is crucial for reliable experimental results. Validate your EDIL3 antibody through:

• Competing peptide assays:

  • Pre-incubate the antibody with the immunizing peptide

  • Compare staining patterns between blocked and unblocked antibody

  • Specific signals should be significantly reduced or eliminated

• Knockout/knockdown controls:

  • Use EDIL3 knockout tissues/cells or siRNA-mediated knockdown

  • Compare staining between wild-type and knockout/knockdown samples

  • Specific antibodies should show reduced or absent signal in knockout/knockdown samples

• Multiple antibody approach:

  • Use antibodies targeting different epitopes of EDIL3

  • Concordant results from different antibodies support specificity

  • For example, compare results from N-terminal antibodies (ABIN2784767) with those targeting other regions

• Recombinant protein controls:

  • Test antibody against purified recombinant EDIL3

  • Confirm single band detection at the expected molecular weight

  • MAB6046 has been validated against recombinant human EDIL3 in direct ELISAs and Western blots

• Cross-reactivity testing:

  • Test against closely related proteins if available

  • MAB6046 showed no cross-reactivity with recombinant human MGF-E8

How can I study EDIL3's role in cancer progression using antibody-based approaches?

EDIL3 has been implicated in various cancer types with context-dependent functions. Here are methodological approaches using antibodies:

• Expression profiling across cancer stages:

  • Use IHC with EDIL3 antibodies on tissue microarrays containing different cancer stages

  • Correlate expression with clinicopathological parameters and patient outcomes

  • Apply digital pathology methods for quantitative assessment

  • Recent studies showed elevated EDIL3 expression in gastric cancer correlating with worse prognosis

• Co-localization studies:

  • Perform dual immunofluorescence with EDIL3 and integrin antibodies (particularly αvβ3 and αvβ5)

  • Use confocal microscopy to assess spatial relationships

  • Analyze co-localization coefficients quantitatively

• Functional analysis in cancer models:

  • Combine antibody-based detection with genetic manipulation (overexpression/knockdown)

  • Assess effects on cancer cell behaviors: migration, invasion, angiogenesis

  • Use antibody-based protein detection methods (Western blot, ELISA) to confirm manipulation success

• Secretome analysis:

  • Use antibody-based techniques to detect EDIL3 in conditioned media

  • Compare secretion levels between normal and cancer cells

  • Correlate with invasive/metastatic potential

• Mechanistic investigations:

  • Employ phospho-specific antibodies against downstream signaling molecules

  • Assess how EDIL3 manipulation affects integrin-mediated signaling pathways

  • Correlate with changes in angiogenic transcription factor expression (e.g., HoxD3)

What are the key considerations when investigating EDIL3 in angiogenesis research?

EDIL3 plays significant roles in angiogenesis through multiple mechanisms. Consider these methodological approaches:

• Endothelial cell models:

  • Use EDIL3 antibodies to track protein expression during angiogenic stimulation

  • Compare expression in quiescent versus activated endothelial cells

  • Research has shown EDIL3 promotes endothelial cell adherence and migration

• Tube formation assays:

  • Apply EDIL3 neutralizing antibodies to inhibit function

  • Quantify effects on endothelial network formation in vitro

  • EDIL3 may inhibit formation of vascular-like structures in some contexts

• Signaling pathway analysis:

  • Use antibodies to detect changes in Bcl-2 expression (EDIL3 upregulates Bcl-2)

  • Assess HoxD3 expression levels, which are increased through EDIL3-αvβ5 integrin binding

  • Monitor αvβ3 integrin and uPA activation status

• In vivo angiogenesis models:

  • Use antibodies to detect EDIL3 in ischemic tissues (muscles, brain, heart)

  • Compare with normal tissues where EDIL3 expression becomes quiescent after birth

  • Correlate with vascular density and blood flow measurements

• Therapeutic investigations:

  • Study effects of exogenous EDIL3 application in ischemia models

  • Use antibodies to track endogenous versus exogenous protein

  • Research has shown beneficial effects in murine models of hind-limb, cardiac, and cerebral ischemia

How do I troubleshoot common problems with EDIL3 antibody experiments?

When facing challenges with EDIL3 antibody-based experiments, consider these methodological solutions:

• Weak or no signal in Western blots:

  • Problem: EDIL3 is primarily secreted; cellular levels may be low

  • Solution: Concentrate cell culture supernatants or use secretion inhibitors

  • Problem: Antibody epitope may be masked by protein folding or modifications

  • Solution: Try different antibodies targeting various EDIL3 regions

• Inconsistent staining in IHC:

  • Problem: Variability in tissue fixation affecting epitope accessibility

  • Solution: Optimize antigen retrieval methods (try both citrate and EDTA buffers)

  • Problem: Batch-to-batch antibody variation

  • Solution: Use antibodies with consistent production methods, preferably monoclonal

• Cross-reactivity issues:

  • Problem: Antibody recognizing related proteins

  • Solution: Validate with blocking peptides and in samples lacking EDIL3

  • Problem: Non-specific binding in certain tissues

  • Solution: Optimize blocking conditions and consider more stringent washing

• Discrepant results between applications:

  • Problem: Different antibody performance across applications

  • Solution: Select application-validated antibodies (e.g., MAB6046 for ELISAs, ABIN2784767 for WB)

  • Problem: Epitope availability differs between native and denatured conditions

  • Solution: Use conformation-specific antibodies matched to your application

• Conflicting experimental outcomes:

  • Problem: EDIL3 functions differ between tissue/cell types

  • Solution: Carefully control for cell type-specific effects

  • Problem: Context-dependent protein interactions

  • Solution: Use co-immunoprecipitation to verify interaction partners in your specific model

How should I interpret different EDIL3 expression patterns across tissue types?

EDIL3 expression varies significantly across tissues and disease states, requiring careful interpretation:

• Developmental context:

  • EDIL3 is actively expressed during embryonic vascular development

  • Expression becomes quiescent in most normal adult tissues

  • Reactivation occurs in specific pathological conditions

• Cancer tissue interpretation:

  • Elevated expression observed in multiple cancer types, including gastric, hepatocellular, pancreatic, colorectal, and breast cancer

  • Decreased expression reported in non-small cell lung cancer

  • Consider both tumor cells and tumor microenvironment (particularly endothelial cells)

  • Correlate with invasion depth, metastatic status, and patient outcomes

• Vascular pathology:

  • Re-expression occurs in ischemic tissues (muscles, brain, heart)

  • May indicate ongoing angiogenic processes

  • Consider co-analysis with other angiogenic markers

• Quantification approaches:

  • Use digital pathology tools for objective quantification of IHC staining

  • Consider both staining intensity and percentage of positive cells

  • Develop standardized scoring systems for cross-study comparability

• Subcellular localization:

  • EDIL3 is primarily secreted, so extracellular matrix localization is expected

  • Intracellular staining may represent newly synthesized protein

  • Abnormal localization patterns may have functional implications

What are the key challenges in correlating EDIL3 expression with clinical outcomes?

Researchers face several methodological challenges when attempting to establish EDIL3 as a prognostic biomarker:

• Antibody selection effects:

  • Different antibodies may detect different isoforms or modified forms of EDIL3

  • Epitope accessibility varies between tissue preparation methods

  • Standardize antibody selection across cohorts for valid comparisons

• Quantification methodology:

  • Subjective scoring systems introduce inter-observer variability

  • Digital pathology approaches require validation

  • Develop clear thresholds for "high" versus "low" expression

• Contextual interpretation:

  • EDIL3 functions differently across cancer types

  • Expression in tumor cells versus stromal cells may have distinct implications

  • Consider integrated analysis with other biomarkers

• Sample considerations:

  • Tissue heterogeneity affects expression assessment

  • Small biopsies may not represent the entire tumor

  • Consider multiple sampling or tissue microarray approaches

• Validation requirements:

  • Preliminary findings require independent cohort validation

  • Multi-institutional studies strengthen evidence

  • Meta-analysis approaches can help resolve conflicting results

How can EDIL3 antibodies contribute to developing novel therapeutic approaches?

EDIL3 antibodies can facilitate therapeutic research through several methodological approaches:

• Target validation studies:

  • Use neutralizing antibodies to block EDIL3 function in preclinical models

  • Assess effects on tumor growth, angiogenesis, and metastasis

  • Determine context-specific therapeutic potential

• Biomarker development:

  • Establish standardized EDIL3 detection protocols for patient stratification

  • Correlate expression with treatment response

  • Develop companion diagnostic approaches

• Monitoring therapeutic response:

  • Track EDIL3 expression changes during treatment

  • Correlate with clinical outcomes

  • Establish whether EDIL3 serves as a resistance mechanism

• Therapeutic antibody development:

  • Use research antibodies to identify optimal epitopes for therapeutic targeting

  • Develop humanized antibodies against EDIL3 for clinical testing

  • Explore antibody-drug conjugate approaches

• Combination therapy research:

  • Investigate EDIL3 targeting in combination with:

    • Anti-angiogenic therapies (given EDIL3's role in angiogenesis)

    • Immune checkpoint inhibitors (considering EDIL3's immunomodulatory functions)

    • Conventional chemotherapy or radiation

What emerging technologies will enhance EDIL3 antibody applications in research?

Future EDIL3 research will benefit from several technological advances:

• Single-cell analysis approaches:

  • Single-cell Western blotting for heterogeneity assessment

  • Imaging mass cytometry for spatial protein expression analysis

  • Single-cell secretome analysis for EDIL3 secretion patterns

• Advanced imaging techniques:

  • Super-resolution microscopy for detailed localization studies

  • Intravital imaging to track EDIL3 dynamics in vivo

  • Correlative light and electron microscopy for ultrastructural context

• Proximity labeling approaches:

  • BioID or APEX2 fusions with EDIL3 to identify proximal interacting proteins

  • Combine with antibody-based validation of novel interactions

  • Map the EDIL3 interactome in different cellular contexts

• Spatial transcriptomics integration:

  • Correlate EDIL3 protein expression with spatial gene expression patterns

  • Identify co-regulated genes in specific tissue microenvironments

  • Develop integrated multi-omic analysis approaches

• Antibody engineering advances:

  • Development of recombinant antibody fragments for improved tissue penetration

  • Bispecific antibodies targeting EDIL3 and its receptors simultaneously

  • Genetically encoded intrabodies for live-cell EDIL3 tracking

How might EDIL3 antibodies be applied in emerging research areas?

EDIL3 antibody applications are expanding into several cutting-edge research domains:

• Liquid biopsy development:

  • Detection of circulating EDIL3 in patient serum as a non-invasive biomarker

  • Correlation with disease progression and treatment response

  • Combination with other circulating biomarkers for improved sensitivity

• Extracellular vesicle research:

  • Investigation of EDIL3 incorporation into exosomes and microvesicles

  • Analysis of vesicle-mediated EDIL3 transfer between cells

  • Functional consequences in recipient cells

• Tissue engineering applications:

  • Monitoring EDIL3 expression during vascular network formation

  • Manipulation of EDIL3 levels to enhance vascularization of engineered tissues

  • Development of EDIL3-functionalized biomaterials

• Immunomodulation research:

  • Exploring EDIL3's role in leukocyte-endothelial adhesion

  • Analysis of EDIL3-mediated inflammatory cell recruitment inhibition

  • Potential applications in inflammatory disease models

• Developmental biology:

  • Investigation of EDIL3's role in embryonic vascular development

  • Application in stem cell differentiation monitoring

  • Potential connections to developmental disorders affecting vasculature