Anti-E Cadherin抗体[M168] (ab76055)

概述

  • 产品名称Anti-E Cadherin抗体[M168]
    参阅全部 E Cadherin 一抗
  • 描述
    小鼠单克隆抗体[M168] to E Cadherin
  • 特异性Ab76055 does not crossreact 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.
  • 经测试应用适用于: Flow Cyt, ICC/IF, IHC-P, IHC-Fr, WB, IP, ELISA, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Horse, Human
  • 免疫原

    Recombinant fragment containing amino acids in the C terminal region of Mouse E Cadherin. This sequence is highly conserved in human and rat E Cadherin.

  • 阳性对照
    • WB: Human A431 cells treated with pervanadate (1 mM) for 30 min. IHC-P: human normal colon tissue sections IF/ICC: A431 cell line

性能

应用

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.

应用 Ab评论 说明
Flow Cyt 1/100.

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.

IP 1/100.
ELISA 1/2000.
ICC 1/250.

靶标

  • 功能Cadherins 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.
  • 组织特异性Non-neural epithelial tissues.
  • 疾病相关Defects 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.
  • 序列相似性Contains 5 cadherin domains.
  • 翻译后修饰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 disassembly of adherens junctions.
  • 细胞定位Cell 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
  • 数据库链接
  • 别名
    • Arc 1 antibody
    • CADH1_HUMAN antibody
    • Cadherin 1 antibody
    • cadherin 1 type 1 E-cadherin antibody
    • Cadherin1 antibody
    • CAM 120/80 antibody
    • CD 324 antibody
    • CD324 antibody
    • CD324 antigen antibody
    • cdh1 antibody
    • CDHE antibody
    • E-Cad/CTF3 antibody
    • E-cadherin antibody
    • ECAD antibody
    • Epithelial cadherin antibody
    • epithelial calcium dependant adhesion protein antibody
    • LCAM antibody
    • Liver cell adhesion molecule antibody
    • UVO antibody
    • Uvomorulin antibody
    see all

Anti-E Cadherin antibody [M168] 图像

  • All lanes : Anti-E Cadherin antibody [M168] (ab76055) at 1/1000 dilution

    Lane 1 : Human A431 cells treated with pervanadate (1 mM) for 30 min
    Lane 2 : Human A431 cells treated with pervanadate (1 mM) for 30 min, then treated with akaline phosphatase


    Predicted band size : 97 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)
    Membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
  • IHC image of E Cadherin staining in human normal 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 (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab76055, 5µg/ml, for 15 mins 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 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise 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 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing A431 cells stained with ab76055 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. 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 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/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 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • 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.

    See Abreview

Anti-E Cadherin antibody [M168] (ab76055)参考文献

This product has been referenced in:
  • Li C  et al. High-Content Functional Screening of AEG-1 and AKR1C2 for the Promotion of Metastasis in Liver Cancer. J Biomol Screen 21:101-7 (2016). Read more (PubMed: 26318406) »
  • Lu Y  et al. MiR-29c inhibits cell growth, invasion, and migration of pancreatic cancer by targeting ITGB1. Onco Targets Ther 9:99-109 (2016). WB ; Human . Read more (PubMed: 26766915) »

See all 13 Publications for this product

Product Wall

The antibody ab76055 is provided at 0.25 mg/mL concentration.

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Sample Human Cell (mda mb 468 breast adenocarcinoma)
Specification mda mb 468 breast adenocarcinoma
Permeabilization No
Fixative Formaldehyde
Username

Mr. ELI Raveh

Verified customer

提交于 Jan 16 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Jaculus jaculus Tissue sections (intestine)
Antigen retrieval step Enzymatic - Buffer/Enzyme Used: ProK
Permeabilization Yes - 0.1% Triton
Specification intestine
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Mai Tran

Verified customer

提交于 Oct 28 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Kidney)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization Yes - 0.1% Tween 20 in PBS
Specification Kidney
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Ms. Chia-Li Lin

Verified customer

提交于 Jan 07 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Lung)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization Yes - 0.1% Tween 20 in PBS
Specification Lung
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Ms. Chia-Li Lin

Verified customer

提交于 Nov 26 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Lung)
Permeabilization Yes - 0.1% Tween 20 in PBS
Specification Lung
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Ms. Chia-Li Lin

Verified customer

提交于 Nov 20 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (intestine)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH9 (Dako)
Specification intestine
Blocking step commercial (Dako) as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 03 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Permeabilization Yes - 0.5% TritonX100 in PBS
Specification HeLa
Fixative Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

提交于 Aug 07 2015

This E-cadherin antibody detects a 120 kDa protein in human A431 cells, and does not cross-react with VE-cadherin or N-cadherin.

We have not tested the cross-reactivity with P-cadherin however according to the sequence homology, there is 80% h...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer pH6
Sample Mouse Tissue (intestine)
Specification intestine
Blocking step goat Fab anti mouse IgG as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jun 13 2014

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