Overview

  • Product name
    Anti-E Cadherin antibody [EPR699]
    See all E Cadherin primary antibodies
  • Description
    Rabbit monoclonal [EPR699] to E Cadherin
  • Host species
    Rabbit
  • Specificity
    E-cadherin contains a number of cleavage sites which may yield a complex fragmentation pattern in WB. Multiple bands between ~80-120 kDa may be observed.
  • Tested applications
    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human E Cadherin aa 150-250.
    Database link: P12830

  • Positive control
    • WB: A431, MCF7, Caco-2, HeLa whole cell lysate (ab150035).
  • General notes

    Mouse, Rat, We have preliminary internal testing data sugesting that 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 ab133597 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 1/1000 - 1/10000. Detects a band of approximately 80-120 kDa (predicted molecular weight: 97 kDa).
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.
  • 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

    • Anti-E Cadherin antibody [EPR699] (ab133597) at 1/1000 dilution (Purified) + MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 97 kDa
      Observed band size: 120 kDa
      why is the actual band size different from the predicted?

    • Lane 1 : Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/5000 dilution
      Lane 2 : Anti-E Cadherin antibody [EPR699] (ab133597) at 1/2000 dilution
      Lane 3 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

      All lanes : PC-3 (Human prostate adenocarcinoma epithelial cell) Whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

      Predicted band size: 97 kDa



      Exposure time: 3 minutes for Ab40772 and Ab133597, 32 seconds for GAPDH

      Blocking and diluting buffer: 5% NFDM/TBST

      Multi-bands can refer to PMID: 11212238; PMID: 14695147 and PMID: 22659456

      This image was produced using unpurified antibody.

    • Lane 1 : Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/5000 dilution
      Lane 2 : Anti-E Cadherin antibody [EPR699] (ab133597) at 1/2000 dilution
      Lane 3 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

      All lanes : HT-29 (Human colorectal adenocarcinoma epithelial cell). Whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 97 kDa
      Observed band size: 80 kDa why is the actual band size different from the predicted?



      Exposure time: 1 second for Ab40772, 3 minutes for Ab133597, 32 seconds for GAPDH.

      Blocking and diluting buffer: 5% NFDM/TBST

      Multi-bands can refer to PMID: 11212238; PMID: 14695147 and PMID: 22659456

      This image was produced using unpurified antibody.

    • All lanes : Anti-E Cadherin antibody [EPR699] (ab133597) at 1/1000 dilution

      Lane 1 : A431 (Human epidermoid carcinoma cell line) cell lysate
      Lane 2 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
      Lane 3 : Caco-2 (Human colorectal adenocarcinoma cell line) cell lysate
      Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 97 kDa



      This image was produced using unpurified antibody.

    References

    This product has been referenced in:
    • Wu X  et al. Overexpression of MCPH1 inhibits the migration and invasion of lung cancer cells. Onco Targets Ther 11:3111-3117 (2018). Read more (PubMed: 29872322) »
    • Cao Y  et al. LncRNA ATB promotes proliferation and metastasis in A549 cells by down-regulation of microRNA-494. J Cell Biochem 119:6935-6942 (2018). Read more (PubMed: 29693289) »
    See all 20 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Question
    Answer

    Thank you for contacting us. I am sorry your customer is experiencing difficulty with this product. A couple of tips they can try to improve the image.

    1) Reduce blocking step to 30 min

    2) Reduce washes to 3 x min min

    3) Check that the secondary antibody is working with other primaries.

    4) Check by Comassie stain to see the protein was tranferred properly. E cadherin is much higher MW than actin, so this would be to be sure the higher MW proteins tranferred well.

    Please let me know if these tips do not improve your customer's results.

    Read More

    Answer

    Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

    Having reviewed the protocol details, I believe this product should have given satisfactory results. I am sorry that this antibody did not perform as stated on the datasheet. After having done a bit of research, I believe that the band at 150 kDa is the band of interest. E-cadherin is synthesized as a 140 kDa precursor that undergoes cleavage by 68 proprotein convertase family proteins associated with the trans-Golgi network (TGN) that 69 remove an amino-terminal pro-region (Posthaus et al., 1998; Beavon, 2000) and I believe that it is this precursor that is detected at 150 kDa. Other publications have observed degradation fragments at 80 kDa (Clin Cancer Res. 2003 Dec 15;9(17):6447-52),60, 45, 38, 33, 29 kDa which could help explain some of the bands on your western blot.

    As requested I have asked our Finance department to issue a credit note for you

    Your credit note ID is xxxxxxxxxxx

    The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

    I apologize for the inconvenience and am pleased to offer you this refund in compensation.

    Thank you for your cooperation.

    Read More

    Answer

    Thank you for contacting us.

    I am sorry to hear you are experiencing difficulties with ab133597. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. If you could also supply us with an image, this will further help us to investigate this case internally and initiate additional testing where necessary.

    If the product was purchased in the last six months and is being used according to our Abpromise, I would be happy to refund this antibody for you.

    I look forward to receiving your reply.

    Read More

    Answer

    Thank you for contacting us.
    The immunogen of ab53033 corresponds to the internal sequence of E-cadherin so this may not suitable as per your requirements.
    The immunogens of antibodies ab15148, ab77287, ab128804, and ab133597 are located within the extra cellular domain of E-cadherin so these antibodies might be suitable. ab133597 has been tested in ICC/IF with mouse and rat cells so it is the most suitable candidate. However the other antibodies can be offered with either mouse or rat or with ICC/IF 100% Abreview discount.
    The Abpromise guarantee is with ICC/IF when performed on fixed cells we do not know which antibody would be suitable for live cell imaging so the suitability has t o be determined by the end user.
    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

    Sign up