Overview

  • Product name
    Anti-E Cadherin antibody [M168] - C-terminal
    See all E Cadherin primary antibodies
  • Description
    Mouse monoclonal [M168] to E Cadherin - C-terminal
  • Host species
    Mouse
  • Specificity
    ab76055 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 applications
    Suitable for: Flow Cyt, ICC/IF, IHC-P, IHC-Fr, WB, IP, ELISA, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Horse, Human
  • Immunogen

    Recombinant fragment corresponding to Mouse E Cadherin (C terminal).

  • Positive control
    • WB: Human A431 cells treated with pervanadate (1 mM) for 30 minutes. IHC-P: Human colon tissue. IF/ICC: A431 cells.

Properties

Applications

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.

Application Abreviews Notes
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.

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

Images

  • 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.

    See Abreview

  • 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.

References

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) »
See all 115 Publications for this product

Customer reviews and Q&As

1-10 of 23 Abreviews or Q&A

Question
Answer

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

Read More
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

Mr. Eli Raveh

Verified customer

Submitted Jan 16 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Catshark Tissue sections (Whole embryo section)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: OmniPrep (Zytomed)
Permeabilization
Yes - TritonX100 0.25% or Tween20 0.1%
Specification
Whole embryo section
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Rami Reshef

Verified customer

Submitted Mar 18 2019

Application
Western blot
Sample
Chicken Cell lysate - whole cell (Chicken gut crypt cells)
Gel Running Conditions
Reduced Denaturing (4-12% SDS PAGE)
Loading amount
50 µg
Specification
Chicken gut crypt cells
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Mar 15 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Tammar wallaby Tissue sections (Testis- Day 1 pouch young and Day 10 PY)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris
Permeabilization
No
Specification
Testis- Day 1 pouch young and Day 10 PY
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 03 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6
Permeabilization
No
Specification
Liver
Blocking step
Protein Block serum free from Dako as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 29 2017

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

Dr. Ann Wheeler

Verified customer

Submitted May 17 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 10 2017

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

Mai Tran

Verified customer

Submitted 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

Miss. Chia-Li Lin

Verified customer

Submitted Jan 07 2016

1-10 of 23 Abreviews or Q&A

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