Overview

  • Product name
  • Description
    Rabbit polyclonal to E Cadherin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human, Pig
  • Immunogen

    Recombinant fragment within Human E Cadherin aa 600-750. The exact sequence is proprietary.
    Database link: P12830

  • Positive control
    • Skin

Properties

Applications

Our Abpromise guarantee covers the use of ab15148 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
WB 1/500. Detects a band of approximately 120 kDa.
ICC/IF Use at an assay dependent concentration.
IHC-P 1/30. for 10 min at RT. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.
IHC-Fr 1/50.

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

  • All lanes : Anti-E Cadherin antibody (ab15148) at 1/500 dilution (for 16 hours at 4°C)

    All lanes : Human OE33 cell - whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/1000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 120 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking Step: 5% BSA fro 1 hour at 23°C

    See Abreview

  • ab15148 staining E Cadherin in Human breast cancer MDA-MB-231 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were treated with ehanol (vehicle) as control or Origanummarjorana extract for 24 hours. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS and blocked with 5% milk for 30 minutes at room temperature. Samples were incubated with primary antibody overnight at 4°C. An Alexa Fluor 488-conjugated Goat anti-rabbit IgG (H+L) polyclonal (1/200) was used as the secondary antibody.

  • ab15148 staining human MCF10A cells by ICC/IF.  Cells were fixed with paraformaldehyde and blocked using 10% serum for 30 minutes at 25 °C.  The primary antibody was diluted 1/25 in TBST and incubated for 1 hour at 25 °C.  An Alexa Fluor® 555 goat anti-rabbit was used as the secondary antibody.

    See Abreview

  • ab15148 staining E Cadherin in Human AGS Gastric Carcinoma tissue sections by Immunohistochemistry (Frozen sections). The sections were acetone fixed and blocked in 5% serum for 1 hour at 23°C. The primary antibody was diluted 1/50 in blocking buffer and incubated with the sample for 1 hours at 23°C.  An HRP-conjugated Goat polyclonal to Rabbit IgG, diluted 1/200, was used as the secondary.

    See Abreview

  • ab15148 staining E Cadherin in Pig Cervix uteri tissue sections by IHC-P (Formaldehyde-fixed, Paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with 10% goat serum for 1 hour at 37°C; antigen retrieval was by heat mediation in 10mM citrate at pH 6 for 2 minutes. The sample was incubated with primary antibody (1/50) at 4°C for 12 hours. An HRP-conjugated goat polyclonal to rabbit IgG (undiluted) was used as secondary antibody.

    See Abreview

  • Immunohistochemical staining of formalin fixed paraffin embedded human skin using ab15148.
  • ab15148 staining E Cadherin in human stomach tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citrate pH 6.0 and then blocking with 5% serum for 1 hour at 23°C was performed. The primary antibody was used diluted 1/50 and incubated with sample for 1 hour at 23°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary antibody.

    See Abreview

  • ab15148 staining E cadherin in Human AGS Gastric carcinoma cultured cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilzed with 0.025% Trton X-100 in TBS and blocked with 5% serum for 1 hour at 23°C. Samples were incubated with primary antibody (1/50 in blocking buffer) for 1 hour at 23°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Ding Y  et al. Megakaryocytic leukemia 1 (MKL1) mediates high glucose induced epithelial-mesenchymal transition by activating LOX transcription. Biochem Biophys Res Commun 509:633-640 (2019). Read more (PubMed: 30553442) »
  • Yang X & Meng T MicroRNA-431 affects trophoblast migration and invasion by targeting ZEB1 in preeclampsia. Gene 683:225-232 (2019). Read more (PubMed: 30315928) »
See all 93 Publications for this product

Customer reviews and Q&As

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1-10 of 13 Abreviews

Application
Western blot
Sample
Dog Cell lysate - other (MDCK cells)
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Loading amount
10 µg
Specification
MDCK cells
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jan 03 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (Swine testicle cells)
Permeabilization
Yes - Triton, 0.1%, 2min
Specification
Swine testicle cells
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 15 2015

Application
ELISA
Sample
Human Cell (H9 embryonic stem cells)
Specification
H9 embryonic stem cells
Type
Indirect
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 11 2013

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (H9 embryonic stem cells)
Specification
H9 embryonic stem cells
Permeabilization
Yes - 0.01% Triton X100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Oct 04 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (MCF-7)
Loading amount
100000 cells
Specification
MCF-7
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (OE33 cells)
Loading amount
20 µg
Specification
OE33 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 27 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (AGS Gastric Carcinoma cells)
Specification
AGS Gastric Carcinoma cells
Fixative
Acetone
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted May 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (AGS Gastric Carcinoma cells)
Specification
AGS Gastric Carcinoma cells
Fixative
Formaldehyde
Permeabilization
Yes - 0.025% Triton-X in TBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Apr 19 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (AGS)
Specification
AGS
Fixative
Formaldehyde
Permeabilization
Yes - 0.025% Triton-X in TBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Feb 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Cervix uteri)
Specification
Cervix uteri
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate, pH 6, 2 min
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Feb 18 2010

1-10 of 13 Abreviews

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