CDH1 Monoclonal Antibody
Description
Table 1: Key Features of Select CDH1 Monoclonal Antibodies
Applications and Performance
These antibodies are versatile across multiple laboratory techniques:
Table 2: Recommended Dilutions and Applications
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Immunohistochemistry: AE00264 demonstrates plasma membrane staining in breast carcinoma at 0.5–1 µg/ml with pH6 epitope retrieval .
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Western Blot: Clone 9C8 detects CDH1 at 97.5 kDa, consistent with its molecular weight .
Specificity and Validation
Rigorous validation ensures minimal cross-reactivity:
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AE00264: Tested against cadherins 2/3 on protein arrays; no cross-reactivity observed (Z-score differential >29) .
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Cusabio Antibodies: Verified via ELISA, WB, IF, and FC, confirming specificity for human CDH1 .
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Bioss 9C8: Recognizes CDH1 in human samples only, with no reactivity to murine or rat homologs .
Biological Relevance of CDH1
CDH1 plays pivotal roles in:
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Cell Adhesion: Maintains epithelial integrity by mediating calcium-dependent homophilic binding .
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Cancer Progression: Loss of CDH1 expression correlates with tumor invasiveness in breast, gastric, and colorectal cancers .
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Therapeutic Targets: CDH1 degradation products (e.g., CTF2) inhibit amyloidogenic pathways, suggesting roles in neurodegenerative disease research .
Research Advancements
Recent studies highlight:
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
The CDH1 monoclonal antibody is a highly specific and sensitive reagent for detecting human CDH1 expression in biological samples. Its production is based on hybridoma technology. The recombinant human CDH1 protein (amino acids 155-707) is used to immunize mice to isolate spleen cells, which are then fused with myeloma cells to generate hybridomas. Hybridomas producing the CDH1 antibody are selected, cultured, and harvested for the desired antibodies. After protein G purification, this CDH1 monoclonal antibody achieves a purity exceeding 95%. Its specificity has been validated through four applications: ELISA, Western blotting, immunofluorescence, and flow cytometry.
CDH1 plays a crucial role in cell adhesion and tissue architecture within epithelial tissues. The CDH1 protein regulates various cellular processes including cell differentiation, proliferation, and apoptosis. It also plays a vital role in maintaining the polarity and integrity of epithelial tissues.
Target Background
- Pathogenic variants are described in four genes encoding components of the p120-catenin complex (CTNND1, PLEKHA7, PLEKHA5) and an epithelial splicing regulator (ESRP2), in addition to the known Cleft lip/Palate-associated gene, CDH1, which encodes E-cadherin. PMID: 29805042
- NEDD9, E-cadherin, and gamma-catenin proteins play roles in pancreatic ductal adenocarcinoma. PMID: 29924959
- Detection of Ezrin and E-cadherin expression in cervical smears may serve as a potential prognostic marker for identifying cervical lesions with a high risk of progression to invasive cervical cancer. This may aid in selecting appropriate therapy or avoiding unnecessary treatment. PMID: 29587669
- CDH1 plays an essential role in epithelial cell adherence. Mutations in CDH1 that cause blepharocheilodontic syndrome impair the cell adhesion function of the cadherin-catenin complex in a dominant-negative manner. PMID: 29348693
- These data suggest that the S18-2 protein induces epithelial to mesenchymal cell transition through the TWIST2/E-cadherin signaling pathway, consequently leading to CXCR4-mediated migration of prostate cancer cells. PMID: 29396484
- The current study demonstrated that miR711-mediated downregulation of CD44 expression inhibited EMT of gastric cancer cells in vitro and in vivo by downregulating vimentin protein expression and upregulating E-cadherin protein expression, as confirmed through transfection, qRT-PCR, and western blotting. PMID: 30226620
- Soluble E-cadherin (sE-cad) (an 80-kDa soluble form), which is highly expressed in the malignant ascites of ovarian cancer patients, is a potent inducer of angiogenesis. In addition to ectodomain shedding, we provide further evidence that sE-cad is abundantly released in the form of exosomes. PMID: 29891938
- In the former, p53 binds to the CDH1 (encoding E-cadherin) locus to antagonize EZH2-mediated H3K27 trimethylation (H3K27me3) to maintain high levels of acetylation of H3K27 (H3K27ac). PMID: 29371630
- E-cadherin silencing relies on the formation of a complex between the paRNA and microRNA-guided Argonaute 1 that, together, recruit SUV39H1 and induce repressive chromatin modifications in the gene promoter. PMID: 28555645
- These results illustrate how E-cadherin instructs the assembly of the LGN/NuMA complex at cell-cell contacts, and define a mechanism that couples cell division orientation to intercellular adhesion. PMID: 28045117
- Low CDH1 expression is associated with pancreatic cancer. PMID: 29956814
- The dysregulation of the TET2/E-cadherin/beta-catenin regulatory loop is a critical oncogenic event in HCC progression. PMID: 29331390
- At the molecular level, transcription of the adherens junction protein E-cadherin is upregulated upon nicotinic acid addition, leading to an accumulation of E-cadherin protein at the cell-cell boundary. This can be attributed to nicotinic acid's ability to facilitate the ubiquitination and degradation of Snail1, a transcription factor that represses E-cadherin expression. PMID: 28256591
- Down-regulation of USP48 increases E-cadherin expression and epithelial barrier integrity through reducing TRAF2 stability. PMID: 28874458
- AnxA5 2D-network mediates E-cadherin mobility in the plasmalemma, triggering human trophoblasts aggregation and subsequent cell fusion. PMID: 28176826
- The disassociation of the beta-catenin/E-cadherin complex in the osteoblast membrane under stretch loading and the subsequent translocation of beta-catenin into the nucleus may represent an intrinsic mechanical signal transduction mechanism. PMID: 29901167
- The presence of E-cadherin decreases cortical contractility during mitosis through a signaling cascade leading to multipolar divisions, and its knockout promotes clustering and survival of cells with multiple centrosomes. PMID: 29133484
- E-cadherin expression is not significantly linked to metastatic disease in pancreatic ductal adenocarcinoma. PMID: 29355490
- High CDH1 expression is associated with the Pathogenesis of Adamantinomatous Craniopharyngiomas. PMID: 29625497
- This study provides evidence for genetic polymorphisms of the adherent junction component cadherin gene and the association of its haplotypes with leukoaraiosis. PMID: 30017735
- This research found that E-cadherin, N-cadherin, and fibronectin are involved in CHD4-mediated epithelial-mesenchymal transition. PMID: 29305962
- Up-regulation of H19 in bladder cancer tissues is correlated with clinical stage or metastasis of cancer. By cell transfection to suppress H19 expression in bladder cancer cells, E-cadherin expression is up-regulated, thus weakening metastatic potency of cancer cells. PMID: 29614625
- These findings suggest that PHF8 plays an oncogenic role in facilitating FIP200-dependent autophagic degradation of E-cadherin, EMT, and metastasis in hepatocellular carcinoma (HCC). PHF8 might be a promising target for prevention, treatment, and prognostic prediction of HCC. PMID: 30180906
- When ANXA5 expression increased, cell proliferation was inhibited by regulating the expression of bcl-2 and bax while cell metastasis was suppressed by regulating E-cadherin and MMP-9 expression. PMID: 30010106
- Six2 is negatively correlated with good prognosis and decreases 5-FU sensitivity via suppressing E-cadherin expression in HCC cells. PMID: 29772441
- The -73A > C CDH1 promoter variation may lead to differences in the overall survival of sporadic gastric carcinoma patients and allele-specific repressions of CDH1. PMID: 29168119
- Overexpression of KLF6-SV1 is associated with young patients, and loss of E-cadherin suggests that this variant correlated with the aggressiveness of nasopharyngeal carcinoma. PMID: 29854578
- Smad4 could be considered a central component of EMT transition in human colorectal cancer that combines with transcriptional factors to reduce E-cadherin and alter the expression of the epithelial phenotype. PMID: 29468299
- hnRNP H/F are important for the maintenance and differentiation of embryonic stem cells and this at least in part reflects a switch in TCF3 alternative splicing that leads to repression of CDH1/E-cadherin. PMID: 30115631
- E-Cadherin and epithelial syndecan-1 were more highly expressed in intraluminal/luminal unicystic ameloblastoma than in mural unicystic ameloblastoma and solid/multicystic ameloblastoma, whereas the stromal expression of syndecan-1 was higher in mural unicystic ameloblastoma and solid/multicystic ameloblastoma. PMID: 29850393
- miR-219-5p promotes tumor growth and metastasis of HCC by regulating CDH1 and can serve as a prognostic marker for HCC patients. PMID: 29862272
- Plasma sE-cadherin levels and sE-cadherin/sVE-cadherin ratios are potential biomarkers for COPD. PMID: 29376431
- HDAC inhibitors augmented both E-cadherin and vimentin expression, but their effects varied across different cholangiocarcinoma cell lines. Therefore, the clinical use of HDAC inhibitors in biliary cancer should be considered cautiously. PMID: 29767267
- E-cadherin expression was preserved in 10 (21.28%) of the 47 NSCLCs immunostained with anti-E-cadherin antibody and reduced/absent in 37 of the 47 (78.72%) NSCLCs studied. E-cadherin plays a major role in intercellular adhesion. PMID: 29556623
- CDH1 promoter methylation may be correlated with cervical cancer carcinogenesis, especially for Caucasians. It was associated with histological subtypes. PMID: 29237293
- High UTX expression is independently associated with a better prognosis in patients with esophageal squamous cell carcinoma (ESCC) and downregulation of UTX increases ESCC cell growth and decreases E-cadherin expression. Our results suggest that UTX may be a novel therapeutic target for patients with ESCC. PMID: 29351209
- Data suggest that ECAD, STAT3, Bak, and Bcl-xL are expressed in affected endometrial tissues of women with endometrioid adenocarcinoma depending on neoplasm staging and cell differentiation. This study was conducted using immunohistochemistry of surgically resected tissues. (STAT3 = signal transducer and activator of transcription 3 protein; Bak = pro-apoptotic protein BAK; Bcl-xL = BCL2 associated agonist of cell death) PMID: 28937296
- LncRNA RP11-789C1.1 inhibited EMT in GC through the RP11-789C1.1/miR-5003/E-cadherin axis, which could be a promising therapeutic target for Gastric Cancer. PMID: 29991048
- Using single-molecule localization microscopy, we show that pAJs in these cells reach more than 1 mum in length and consist of several cadherin clusters with crystal-like density interspersed within sparser cadherin regions. Notably, extrajunctional cadherin appears to be monomeric, and its density is almost four orders of magnitude less than observed in the pAJ regions. PMID: 29691319
- CDH1 methylation may play a role in the initiation and progression of salivary carcinoma ex pleomorphic adenoma. PMID: 29207084
- We illustrate the approach using immunohistochemical measurements of the epithelial-mesenchymal transition marker E-cadherin in a set of colorectal primary tumors from a population-based prospective cohort in North Carolina. PMID: 29338703
- The aim of our study was to analyze the immunohistochemical expression of beta-catenin, E-cadherin, and Snail, depending on clinico-morphological aspects of the laryngeal squamous cell carcinomas. Results revealed variable E-cadherin, beta-catenin, and Snail expression, depending on differentiation degree and tumor stage. PMID: 29250652
- Twist, E-cadherin, and N-cadherin protein were differently expressed in endometrioid adenocarcinoma tissues and in normal endometrium which indicates their potential function for endometrioid adenocarcinoma development. PMID: 29237910
- Findings uncover a new regulatory network in RCC involving metastasis-promoting miR-720 that directly targets expression of key metastasis-suppressing proteins E-cadherin and alphaE-catenin complex. PMID: 28802251
- Results show that E-cadherin expression levels were negatively regulated by 90K via ubiquitination-mediated proteasomal degradation in a cell density-dependent manner. PMID: 29207493
- These results indicate that increased alpha-actinin-1 expression destabilizes E-cadherin-based adhesions, which is likely to promote the migratory potential of breast cancer cells. Furthermore, our results identify a-actinin-1 as a candidate prognostic biomarker in basal-like breast cancer. PMID: 29742177
- High glucose enhances the formation of EZH2/Snail/HDAC1 complex in the nucleus, which in turn causes E-cadherin repression. PMID: 29705809
- TGF-beta1 induced epithelial-mesenchymal transition in non-small cell lung cancer cells by upregulating miR-9 and downregulating miR-9's target, E-cadherin. PMID: 29118814
- Results show that the E-cadherin/beta-catenin complex is disrupted by ICAT, promoting epithelial-mesenchymal transition of cervical cancer cells. PMID: 29048651
- Studies categorize cadherin 1 (CDH1) variants as neutral or deleterious. PMID: 29231860
Q&A
What is CDH1 and why is it an important research target?
CDH1 (also known as E-cadherin, Cadherin-1, CD324) is a calcium-dependent cell adhesion glycoprotein that plays crucial roles in epithelial tissue integrity and function. CDH1 is involved in mechanisms regulating cell-cell adhesions, mobility, and proliferation of epithelial cells . It contains five extracellular cadherin repeats, a transmembrane region, and a highly conserved cytoplasmic tail that interacts with catenin proteins to anchor to the actin cytoskeleton .
Research importance of CDH1:
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Functions as a potent tumor suppressor
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Loss of function contributes to cancer progression through increased proliferation, invasion, and metastasis
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Germline pathogenic variants in CDH1 are associated with hereditary diffuse gastric cancer (HDGC) and lobular breast cancer
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Serves as a receptor for bacterial pathogens like Listeria monocytogenes
What applications are CDH1 monoclonal antibodies typically used for?
CDH1 monoclonal antibodies are versatile research tools validated for multiple applications:
How should I select the most appropriate CDH1 monoclonal antibody clone for my research?
Selection of the optimal CDH1 antibody clone depends on several critical factors:
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Epitope recognition: Different clones recognize distinct epitopes. For example, clone CDH1/1525 targets recombinant full-length protein , while clone COH-3 targets a specific region (292-587 aa) . Select based on your research requirements.
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Validated applications: Confirm the antibody is validated for your specific application. For instance:
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Species reactivity: Verify cross-reactivity with your species of interest:
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Publication record: Consider antibodies cited in publications relevant to your research area. For example, some clones like GTX125890 have been cited in 13 publications .
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Subcellular localization requirements: For detecting specific cellular compartments where CDH1 localizes (cell membrane, Golgi apparatus, endosomes) .
What controls should be included when using CDH1 monoclonal antibodies?
Robust experimental design requires appropriate controls:
Positive controls:
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Cell lines with known CDH1 expression (MCF-7 is commonly used)
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Epithelial tissues like normal gastric epithelium or mammary epithelium
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Recombinant CDH1 protein for Western blot
Negative controls:
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CDH1-knockout or knockdown cell lines
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Non-epithelial tissues/cells that don't express CDH1
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Isotype control antibody (matching IgG1 or IgG1κ for most clones)
Additional validation controls:
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For IHC/IF: Omission of primary antibody
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For Western blot: Blocking peptide competition
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For functional studies: Compare multiple antibody clones targeting different epitopes
Normalization controls:
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For Western blot: Housekeeping proteins (β-actin, GAPDH)
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For IHC: Normal adjacent tissue within the same section
What are the optimal sample preparation methods for CDH1 detection in different applications?
For Western Blot:
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Cell lysis: Use RIPA buffer containing protease inhibitors and 1-2 mM calcium to maintain cadherin structure
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Protein amount: Load 20-50 μg of total protein
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Denaturing conditions: Heat samples at 95°C for 5 minutes in sample buffer containing SDS and β-mercaptoethanol
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Gel percentage: Use 7.5-10% SDS-PAGE for optimal separation of the 120-135 kDa CDH1 protein
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Transfer: Use wet transfer method (25V overnight at 4°C) for higher molecular weight proteins like CDH1
For Immunohistochemistry:
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Fixation: 10% neutral buffered formalin for 24-48 hours
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Antigen retrieval: Citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) for 20 minutes
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Blocking: 5% BSA or normal serum (goat/horse) for 1 hour
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Primary antibody incubation: 4°C overnight at dilutions specified by manufacturer (typically 1:50-1:200)
For Flow Cytometry:
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Cell preparation: Single-cell suspension (2-5 × 10^6 cells/mL)
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Fixation: 2% paraformaldehyde for 10-15 minutes (for total protein) or unfixed (for surface expression)
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Permeabilization (if needed): 0.1% Triton X-100 or 0.1% saponin
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Antibody incubation: 30-60 minutes at 4°C
What are common technical issues when using CDH1 antibodies and how can they be resolved?
| Issue | Possible Causes | Solutions |
|---|---|---|
| High background in IHC/IF | Insufficient blocking, antibody concentration too high, non-specific binding | Increase blocking time/concentration, titrate antibody, use more stringent washing, add 0.1-0.3% Triton X-100 to reduce non-specific binding |
| Weak/no signal in Western blot | Protein degradation, inefficient transfer, epitope masking | Include protease inhibitors, ensure calcium in buffers, optimize transfer conditions, try different epitope antibodies |
| Multiple bands in Western blot | Proteolytic cleavage products, glycosylation variants, cross-reactivity | Use fresh samples with protease inhibitors, verify with different antibody clones, include positive controls |
| Membrane vs. cytoplasmic staining variability | Fixation artifacts, epitope accessibility, biological variability in CDH1 localization | Optimize fixation time, use membrane permeabilization for total CDH1, compare with markers of subcellular compartments |
| Inconsistent IHC staining | Tissue fixation variability, antigen retrieval issues | Standardize fixation protocols, optimize antigen retrieval method and time, use automated staining platforms if available |
How can CDH1 monoclonal antibodies be used to study epithelial-mesenchymal transition (EMT) in cancer research?
EMT is a critical process in cancer progression where epithelial cells lose their characteristic features and gain mesenchymal properties. CDH1 downregulation is a hallmark of EMT. Here's a methodological approach to study EMT using CDH1 antibodies:
Experimental design for EMT studies:
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Cell model selection:
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Epithelial cancer cell lines (e.g., MCF-7, A549)
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EMT induction using TGF-β, EGF, hypoxia, or other stimuli
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Time-course experiments (24h, 48h, 72h post-induction)
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Multilevel analysis approaches:
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Western blot: Quantify CDH1 protein level changes relative to mesenchymal markers (N-cadherin, vimentin)
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Immunofluorescence: Visualize CDH1 relocalization from cell junctions to cytoplasm during EMT
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Flow cytometry: Quantify surface CDH1 expression in single cells to identify transitional populations
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Proximity ligation assay: Assess changes in CDH1-catenin interactions during EMT
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Quantification methods:
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Membrane/cytoplasm CDH1 staining ratio
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Co-localization analysis with β-catenin
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Correlation with cell morphology changes
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Validation approaches:
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Correlate CDH1 expression changes with EMT transcription factors (SNAIL, SLUG, ZEB1/2, TWIST)
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Functional migration and invasion assays to correlate with CDH1 loss
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How can CDH1 antibodies be used to differentiate between sporadic and hereditary diffuse gastric cancer (HDGC)?
Hereditary diffuse gastric cancer (HDGC) is associated with germline pathogenic variants in the CDH1 gene. CDH1 antibodies can be used in diagnostic and research settings to evaluate protein expression patterns that might indicate underlying genetic alterations.
Methodological approach:
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Sample collection and preparation:
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FFPE tissue from gastric biopsies or surgical specimens
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Matched normal and tumor tissue from the same patient
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Family history documentation for all cases
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Immunohistochemical analysis:
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Interpretation of staining patterns:
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Sporadic diffuse gastric cancer: Often shows heterogeneous CDH1 loss
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HDGC with CDH1 mutations: Typically shows complete loss of expression in tumor cells with internal positive controls (lymphocytes, stromal cells)
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Quantitative scoring: Percentage of cells with membrane staining and intensity (0-3+)
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Correlation with molecular findings:
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Compare IHC results with CDH1 germline variant status
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In cases with CDH1 germline variants, loss of heterozygosity (LOH) analysis
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Follow-up with second-hit mechanism analysis (promoter hypermethylation, somatic mutations)
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As demonstrated in a study by Benusiglio et al., loss of CDH1 expression in early gastric cancer foci from prophylactic gastrectomy specimens strongly correlates with underlying CDH1 pathogenic variants, providing a useful screening tool before genetic testing .
How should CDH1 staining patterns be interpreted in different tissue and cell types?
Normal epithelial tissues:
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Strong, continuous membrane staining at cell-cell contacts
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Little to no cytoplasmic staining
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Intensity may vary by epithelial type
Cancer tissues:
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Reduced membrane staining
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Increased cytoplasmic localization
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Complete loss in some diffuse/lobular carcinomas
Staining pattern interpretation table:
| Staining Pattern | Biological Interpretation | Technical Considerations |
|---|---|---|
| Strong membrane staining | Normal adherens junction formation | Validate membrane specificity with other markers |
| Reduced/fragmented membrane staining | Partial adherens junction disruption | Distinguish from fixation artifacts |
| Cytoplasmic accumulation | Internalization, trafficking defects | May indicate EMT or post-translational modifications |
| Complete loss | Transcriptional silencing, mutation, or degradation | Verify with alternate antibody clones and controls |
| Nuclear translocation | Rare, may indicate proteolytic processing | Validate with nuclear/cytoplasmic fractionation |
Cell-type specific considerations:
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Epithelial cells: Primary site of expression
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Mesenchymal cells: Typically negative
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Cancer cells: Variable depending on differentiation status
What methodological approaches should be used to quantify CDH1 expression levels in research studies?
Quantification of CDH1 expression should be conducted using standardized approaches to ensure reproducibility:
Western Blot Quantification:
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Use appropriate loading controls (β-actin, GAPDH)
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Employ densitometry software (ImageJ, Image Lab)
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Report relative expression as normalized to control
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Include multiple biological replicates (minimum n=3)
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Statistical analysis: t-test or ANOVA for group comparisons
IHC Quantification Systems:
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H-score: Intensity (0-3+) × percentage of positive cells (0-100%), range 0-300
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Allred score: Intensity score (0-3) + proportion score (0-5), range 0-8
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Membrane completeness: Complete, incomplete, or absent membrane staining
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Digital image analysis: Automated pixel-based quantification systems
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Inter-observer validation: Multiple pathologists scoring the same slides
Flow Cytometry Quantification:
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Mean fluorescence intensity (MFI) normalized to isotype control
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Percentage of CDH1-positive cells in the population
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Generation of standardized MFI using calibration beads
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Correlation with cell size/complexity parameters
Quantitative comparison example table:
