Product nameAnti-E Cadherin antibody [M168] - C-terminal
See all E Cadherin primary antibodies
DescriptionMouse monoclonal [M168] to E Cadherin - C-terminal
Specificityab76055 does not cross react with VE Cadherin or N Cadherin. This product may give a weak signal in Western Blot when using unstimulated cell lines. Abcam recommends using A431 cells treated with pervandate (1mM, 30 minutes) as a positive control. Abcam recommends ab40772 and ab11512 as a alternative when using unstimulated samples in Western Blot.
Tested applicationsSuitable for: Flow Cyt, ICC/IF, IHC-P, IHC-Fr, WB, IP, ELISA, ICCmore details
Species reactivityReacts with: Mouse, Rat, Horse, Human
Recombinant fragment corresponding to Mouse E Cadherin (C terminal).
- WB: Human A431 cells treated with pervanadate (1 mM) for 30 minutes. IHC-P: Human colon tissue. IF/ICC: A431 cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 50% Glycerol, PBS
Concentration information loading...
PurityProtein A purified
Light chain typekappa
Our Abpromise guarantee covers the use of ab76055 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use at an assay dependent concentration. PubMed: 23405249|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|IHC-Fr||Use at an assay dependent concentration. PubMed: 21769484|
|WB||1/100 - 1/1000. Predicted molecular weight: 97 kDa.
This product may give a weak signal in Western Blot when using unstimulated cell lines. Abcam recommends using A431 cells treated with pervandate (1mM, 30 minutes) as a positive control.
FunctionCadherins are calcium-dependent cell adhesion proteins. They preferentially interact with themselves in a homophilic manner in connecting cells; cadherins may thus contribute to the sorting of heterogeneous cell types. CDH1 is involved in mechanisms regulating cell-cell adhesions, mobility and proliferation of epithelial cells. Has a potent invasive suppressor role. It is a ligand for integrin alpha-E/beta-7.
E-Cad/CTF2 promotes non-amyloidogenic degradation of Abeta precursors. Has a strong inhibitory effect on APP C99 and C83 production.
Tissue specificityNon-neural epithelial tissues.
Involvement in diseaseDefects in CDH1 are the cause of hereditary diffuse gastric cancer (HDGC) [MIM:137215]. An autosomal dominant cancer predisposition syndrome with increased susceptibility to diffuse gastric cancer. Diffuse gastric cancer is a malignant disease characterized by poorly differentiated infiltrating lesions resulting in thickening of the stomach. Malignant tumors start in the stomach, can spread to the esophagus or the small intestine, and can extend through the stomach wall to nearby lymph nodes and organs. It also can metastasize to other parts of the body. Note=Heterozygous germline mutations CDH1 are responsible for familial cases of diffuse gastric cancer. Somatic mutations in the has also been found in patients with sporadic diffuse gastric cancer and lobular breast cancer.
Defects in CDH1 are a cause of susceptibility to endometrial cancer (ENDMC) [MIM:608089].
Defects in CDH1 are a cause of susceptibility to ovarian cancer (OC) [MIM:167000]. Ovarian cancer common malignancy originating from ovarian tissue. Although many histologic types of ovarian neoplasms have been described, epithelial ovarian carcinoma is the most common form. Ovarian cancers are often asymptomatic and the recognized signs and symptoms, even of late-stage disease, are vague. Consequently, most patients are diagnosed with advanced disease.
Sequence similaritiesContains 5 cadherin domains.
modificationsDuring 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 disassembly of adherens junctions.
Cellular localizationCell junction. Cell membrane. Endosome. Golgi apparatus > trans-Golgi network. Colocalizes with DLGAP5 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. Colocalizes with RAB11A endosomes during its transport from the Golgi apparatus to the plasma membrane.
- Information by UniProt
- Arc 1 antibody
- CADH1_HUMAN antibody
- Cadherin 1 antibody
Immunohistochemistry staining of E Cadherin in human MMCR (Muller’s muscle conjunctival resection) tissue.
Xylene and rehydration with serial ethanol dilutions were used for deparaffinization. Slides were washed twice for 5 minutes in 0.25% Triton X-100 for permeabilization and blocked for 2 hours at room temperature with 2% BSA and 10% normal donkey serum in PBS. Slides were incubated overnight at 4°C with ab76055. The next day, the slides were washed twice for 5 minutes in PBS and incubated for 1–2 hours with respective secondary antibodies diluted in blocking solution (1:200–800). Vecta shield mounting medium with 4′,6-diamidino-2-phenylindole (DAPI) was placed over the slides and covered with a glass coverslip. Slides were analyzed using the Zeiss LSM 710 Confocal Microscope.
ab76055 staining E Cadherin in mouse intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Tissue was fixed with paraformaldehyde and blocked with 0.1% goat Fab anti-mouse IgG for 30 minutes at 25°C; antigen retrieval was by heat mediation in 10mM citrate buffer, pH 6. Samples were incubated with primary antibody (1/250) for 2 hours at 25°C. An Alexa Fluor® 488-conjugated donkey anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.
Lane 1 : Anti-E Cadherin antibody [M168] - C-terminal (ab76055) at 1/1000 dilution
Lane 2 : Anti-E Cadherin antibody [M168] - C-terminal (ab76055) at 1/2000 dilution
Lane 3 : Anti-E Cadherin antibody [M168] - C-terminal (ab76055) at 1/4000 dilution
Lane 1 : A431 (Human epidermoid carcinoma cell line) denatured whole cell lysate
Lanes 2-3 : A431 denatured whole cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 97 kDa
Exposure time: 2 minutes
Blocked and probed in the presence of 5% milk, and the image was captured using chemiluminescence and film.
IHC image of E Cadherin staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH 6, for 20 minutes. The section was then incubated with ab76055, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab76055 stained A431 (Human epidermoid carcinoma cell line) cells.
The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76055, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
Overlay histogram showing A431 (Human epidermoid carcinoma cell line) cells stained with ab76055 (red line).
The cells were fixed with 4% paraformaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76055, 1/100 dilution) for 30 minutes at 22ºC. The secondary antibody used was a DyLight® 488 goat anti-mouse IgG (H+L) ab96879 at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
This antibody gave a positive signal in A431 cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.
This product has been referenced in:
- Gu TT et al. Pterostilbene alleviates fructose-induced renal fibrosis by suppressing TGF-ß1/TGF-ß type I receptor/Smads signaling in proximal tubular epithelial cells. Eur J Pharmacol 842:70-78 (2019). Read more (PubMed: 30336139) »
- Wang C et al. Upregulation of lncRNA snoRNA host gene 6 regulates NUAK family SnF1-like kinase-1 expression by competitively binding microRNA-125b and interacting with Snail1/2 in bladder cancer. J Cell Biochem 120:357-367 (2019). Read more (PubMed: 30168179) »