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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | FC, WB, FUNC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB3199 |
| Replaces | 04-1103 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-E-Cadherin Antibody, clone 67A4 |
| Alternate Names |
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| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Protein A purified immunoglobulin. Liquid in 0.02M PBS with NaCl, pH=7.6, 0.1% Sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-E-Cadherin Antibody, clone 67A4 detects level of E-Cadherin & has been published & validated for use in FC & WB. |
| Key Applications |
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| Application Notes | Immunoblotting: 123kD in membrane fractions; MCF-7 membranes positive control. Confluent cells were washed three times with ice-cold PBS and lysed in 50mM HEPES pH 7.5, 150mM NaCl, 1mM EDTA, 10% glycerine, 1% triton X-100, 20mM Na(4)P(2)O(7), 2mM Na-vanadate, 100mM NaF, 2mM PMSF, 20μg/mL leupeptin, 1mM pNPP(para-nitrophenylphosphate), 20μg/mL aprotinin, and 200μM NH(4)-molybdate. Crude extracts were clarified by centrifugation (10,000 x g 10 min), and supernatants were taken for immunoprecipitation. Protein concentration of cleared lysates were determined by BCA protein assay. {Buhring, et Al Luekemia 10:106-116, 1996}. FACS Analysis Functional blocking of E-cadherin Does not work well for immunoprecipitation Original paper, Buhring, H et Al Leukemia 10:106-116, 1996. Optimal working dilutions must be determined by end user. |
| Biological Information | |
|---|---|
| Immunogen | T-47D breast carcinoma line |
| Clone | 67A4 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | Antibodies against E-cadherin are suitable to characterize epithelial cells. E-Cadherin is involved in adhesion of epithelial cells (mostly homotypic and homophilic). E-cadherin is also expressed on immature erythroid cells. The antibody 67A4 is suitable to study E-cadherin expression by flow cytometry. In addition, it blocks the interaction of E-cadherin molecules and therefore is suitable to study E-cadherin-mediated signal transduction. Antigen is expressed in non-neural epithelial tissues. |
| Isotype | IgG1 |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This gene is a classical cadherin from the cadherin superfamily. The encoded protein is a calcium dependent cell-cell adhesion glycoprotein comprised of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. Mutations in this gene are correlated with gastric, breast, colorectal, thyroid and ovarian cancer. Loss of function is thought to contribute to progression in cancer by increasing proliferation, invasion, and/or metastasis. The ectodomain of this protein mediates bacterial adhesion to mammalian cells and the cytoplasmic domain is required for internalization. Identified transcript variants arise from mutation at consensus splice sites. |
| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P12830 # E-Cad/CTF2 promotes non-amyloidogenic degradation of Abeta precursors. Has a strong inhibitory effect on APP C99 and C83 production. SIZE: 882 amino acids; 97456 Da SUBUNIT: Homodimer; disulfide-linked. Interacts directly, via the cytoplasmic domain, with CTNNB1 or JUP to form the PSEN1/cadherin/catenin adhesion complex which connects to the actin skeleton through the actin binding of alpha-catenin. Interaction with PSEN1, cleaves CDH1 resulting in the disassociation of cadherin-based adherens junctions (CAJs). Interacts with AJAP1, CTNND1 and DLG7. SUBCELLULAR LOCATION: Cell junction. Cell membrane; Single-pass type I membrane protein. Note=Colocalizes with DLG7 at sites of cell-cell contact in intestinal epithelial cells. Anchored to actin microfilaments through association with alpha-, beta- and gamma-catenin. Sequential proteolysis induced by apoptosis or calcium influx, results in translocation from sites of cell-cell contact to the cytoplasm. TISSUE SPECIFICITY: Non-neural epithelial tissues. PTM: During apoptosis or with calcium influx, cleaved by a membrane-bound metalloproteinase (ADAM10), PS1/gamma-secretase and caspase-3 to produce fragments of about 38 kDa (E-CAD/CTF1), 33 kDa (E-CAD/CTF2) and 29 kDa (E-CAD/CTF3), respectively. Processing by the metalloproteinase, induced by calcium influx, causes disruption of cell-cell adhesion and the subsequent release of beta-catenin into the cytoplasm. The residual membrane-tethered cleavage product is rapidly degraded via an intracellular proteolytic pathway. Cleavage by caspase-3 releases the cytoplasmic tail resulting in disintegration of the actin microfilament system. The gamma-secretase-mediated cleavage promotes disaaaembly of adherens junctions. DISEASE: "SwissProt: P12830 # Defects in CDH1 are involved in dysfunction of the cell- cell adhesion system, triggering cancer invasion (gastric, breast, ovary, endometrium and thyroid) and metastasis. & Defects in CDH1 are a cause of hereditary diffuse gastric cancer (HDGC) [MIM:137215]. & Defects in CDH1 are a cause of susceptibility to endometrial cancer [MIM:608089]." SIMILARITY: SwissProt: P12830 ## Contains 5 cadherin domains. |
| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain 2-8°C in undiluted aliquots for up to 12 months after date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |