Recombinant
RabMAb

Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)

Overview

  • Product name
    Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker
    See all E Cadherin primary antibodies
  • Description
    Rabbit monoclonal [EP700Y] to E Cadherin - Intercellular Junction Marker
  • Specificity
    This antibody has been tested on human samples in both WB and IHC. However,mouse samples have only been tested in WB in the laboratory; customer feedback (see Abreview) suggests the antibody does not perform well in IHC on mouse tissue.
  • Tested applications
    Suitable for: IHC-Fr, ICC/IF, IHC-P, IP, Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to residues in the 5th domain of human E-cadherin

  • Positive control
    • MCF-7 cell lysate; human breast carcinoma; human hepatocellular carcinoma
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab40772 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration. PubMed: 24915897
ICC/IF 1/500.

Permeabilisation is unnecessary as the immunogen is in an extracellular domain.

IHC-P 1/500.
IP 1/50.
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/10000 - 1/50000. Predicted molecular weight: 120 kDa.

Target

  • Function
    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.
  • Tissue specificity
    Non-neural epithelial tissues.
  • Involvement in disease
    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.
  • Sequence similarities
    Contains 5 cadherin domains.
  • Post-translational
    modifications
    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.
  • Cellular localization
    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
  • Database links
  • Alternative names
    • 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 [EP700Y] - Intercellular Junction Marker images

  • Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ab40772. Green-E-Cadherin red-PI

  • ab40772 showing positive staining in human colonic adenocarcinoma tissue.

  • Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/200000 dilution + MCF7 (Human breast adenocarcinoma ) whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) at 1/1000 dilution

    Predicted band size : 120 kDa
    Observed band size : 80,97,100,120 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    Blocking and diluting buffer and concentration 5% NFDM/TBST.

    The full-length of E-cadherin is 120 kDa. The other bands are due to proteolytic cleavages in different Cadherin domains. (Ref: PMID: 14695147)

  • ICC/IF image of ab40772 stained human MCF7 cells. The cells were fixed in Paraformaldehyde, before incubation in 0.1% BSA + 1% goat serum in PBS as a blocking agent for 30 mins at 22oC. The cells were then incubated with the antibody (ab40772, 1:100 dilution) for 2 hours at 22°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG. DAPI was used to stain the cell nuclei (blue).

    See Abreview

  • Overlay histogram showing A431 cells stained with ab40772 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab40772, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • ab40772 showing positive staining in human lung adenocarcinoma tissue.

  • ab40772 showing positive staining in human breast carcinoma tissue.

  • ab40772 showing positive staining in human papillary carcinoma of thyroid gland tissue.

  • ab40772 showing positive staining in human transitional cell carcinoma of kidney tissue.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)

This product has been referenced in:
  • Bi Y  et al. MicroRNA-7 functions as a tumor-suppressor gene by regulating ILF2 in pancreatic carcinoma. Int J Mol Med 39:900-906 (2017). WB ; Human . Read more (PubMed: 28259961) »
  • Huang H  et al. Allicin inhibits tubular epithelial-myofibroblast transdifferentiation under high glucose conditions in vitro. Exp Ther Med 13:254-262 (2017). Human . Read more (PubMed: 28123498) »

See all 54 Publications for this product

Product Wall

Filter by Application

Filter by Species

Filter by Ratings

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Keratinocyte (HaCaT))
Permeabilization
Yes - 0.2% Triton x100
Specification
Keratinocyte (HaCaT)
Fixative
Formaldehyde
Username

Dr. Ann Wheeler

Verified customer

Submitted May 17 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, postnatal day 6)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, postnatal day 6
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Mr. Bryan Niedenberger

Verified customer

Submitted Jul 12 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Pluripotent stem cell)
Permeabilization
Yes - 0.1% Triton X100
Specification
Pluripotent stem cell
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde
Username

Oliver Thompson

Verified customer

Submitted Apr 18 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer, pH6
Sample
Human Tissue sections (colon)
Specification
colon
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 10 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MCF-7)
Specification
MCF-7
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
0.1% BSA + 1% goat serum in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jan 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF-7)
Loading amount
100000 cells
Specification
MCF-7
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jan 28 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (vaginal epithelial cells)
Loading amount
40 µg
Specification
vaginal epithelial cells
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 06 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (DU145 Prostate cancer cell)
Specification
DU145 Prostate cancer cell
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.2% Triton / PBS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 10 2010

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Small Intestine Crypt Epithelia)
Loading amount
15 µg
Specification
Small Intestine Crypt Epithelia
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 30 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (KIDNEY)
Specification
KIDNEY
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 21 2009

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