Overview

  • Product name
    Anti-E Cadherin antibody [DECMA-1]
    See all E Cadherin primary antibodies
  • Description
    Rat monoclonal [DECMA-1] to E Cadherin
  • Host species
    Rat
  • Specificity
    Some of our customers have had good results using ab11512 in human samples, particularly in ICC. In our hands however this product does not work in human samples. Please contact Scientific Support for further information or queries.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Dog
    Predicted to work with: Cow
  • Immunogen

    Mouse embryonal carcinoma cell line PCC4 Aza RI.

  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab11512 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
ICC/IF Use a concentration of 5 - 10 µg/ml.
Flow Cyt Use at an assay dependent concentration. PubMed: 20521328

ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use at an assay dependent concentration. PubMed: 19586906
IHC-Fr Use at an assay dependent concentration.

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

  • ab11512 staining E Cadherin in Mouse embryonic (E14.5) lung tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 1% serum for 3 hours at 4°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 14 hours at 4°C. An Alexa Fluor®488-conjugated Donkey anti-rat IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ab11512 stained in M158 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab11512 at 10 µg/ml overnight at +4°C. The secondary antibody was ab150165 (colored green) used at 1 µg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

  • Sample: Murine derived breast cancer whole cell lysate, 30 µg.

    ab11512 used at a 1/5000 dilution for 16 hours at 4°C.

    Goat polyclonal IRDye 800CW used as the secondary antibody at a 1/10,000 dilution.

     

    See Abreview

  • ab11512 E Cadherin staining canine kidney (MDCK) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 37°C. Samples were incubated with primary antibody, 1/100, in blocking buffer for 1 hour at 37°C. An undiluted Cy3®-conjugated Donkey polyclonal to rat IgG was used as secondary antibody.

    See Abreview

  • ab11512 at 1/500 staining dog kidney cells by ICC/IF. The cells were methanol fixed and blocked with BSA before incubation with the antibody for 18 hours at 4°C. An Alexa Fluor ® 555 conjugated goat anti-rat IgG was used as the secondary.

    See Abreview

References

This product has been referenced in:
  • Smyrek I  et al. E-cadherin, actin, microtubules and FAK dominate different spheroid formation phases and important elements of tissue integrity. Biol Open 8:N/A (2019). Read more (PubMed: 30578251) »
  • Li X  et al. Development of immortalized Hertwig's epithelial root sheath cell lines for cementum and dentin regeneration. Stem Cell Res Ther 10:3 (2019). Read more (PubMed: 30606270) »
See all 63 Publications for this product

Customer reviews and Q&As

1-9 of 9 Q&A

Answer

We have tested the conjugated form of this antibody in flow cytometry, Anti-E Cadherin antibody [DECMA-1] (FITC) (ab150004).

The protocol we used is the following:

The cells were fixed with 80% methanol (5 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 (ab150004, 0.1µg/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was Rat IgG1 [RTK2071] (FITC) (https://www.abcam.com/ab18404.html, 0.1µg/1x106 cells) for 30 min at 22°C. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

The unconjugated form has also been used in FACs in the following two publications which may be of use to you:
http://www.ncbi.nlm.nih.gov/pubmed/20521328?dopt=Abstract
http://www.ncbi.nlm.nih.gov/pubmed/8056851?dopt=Abstract

Looking at customer reviews for this antibody in MDCK cells in ICC, they all included a permeabilisation step, either fixing in methanol or permeabilising in 0.5% Triton X-100. It does seem that in this species, for optimum results, a permeabilisation step should be included. We have no information regarding the results in dog without permeabilisation.

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Answer

Thank you for your phone call.

I am sorry to hear that both antibodies do not work as expected.


As for ab92539, I am sending you - as we discussed - the testing discount code for having tried it in IHC-Fr. After submitting your negative data, you can receive a free antibody of your choice on your next order (please see details below).


As for ab11512, we only have the same lot in stock that you already tried. Please let me know if I shall send you free of charge another vial of the same lot. Other options would be another antibody to the same protein or a different protein, or a credit or refund.

DISCOUNT CODE: xxx for ab92539
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab92539 in IHC-Fr. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for IHC-Frand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you for contacting us. FAM is a fluorochrome with an excitation of 494 and emission wavelength 519. We have tested using secondaries conjugated to Alexa594, Alexa555, and Cy3 with good results.

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

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Answer

Vielen Dank für Ihre Anfrage.

Das Immunogen für ab11512 (Anti-E Cadherin antibody [DECMA-1])n was eine Zellpräparation von einer murinen embryonalen Carcimomzelllinie.

Unseres Wissens wurde das genaue Epitop das der Antikörper bindet nicht genau untersucht.

Vielleicht hilft Ihnen aber folgende Publikation über diesem Antikörperklon weiter:

http://www.ncbi.nlm.nih.gov/pubmed/1710917

Sie zeigt, dass das Epitop in der extrazellulären Domäne nahe der Membran liegt.

Es tut mir leid, dass ich keine weiteren Informationen für Sie haben und hoffe, diese Information ist dennoch hilfreich.

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Answer

Thank you for contacting us.
I have checked the homologies for the mouse and human protein with the zebrafish sequence.
human - zebrafish = 53% (C-terminus aligns better than the rest of the protein)
mouse - zebrafish = 81%
The antibody was raised against mouse embryonal carcinoma cell line PCC4 Aza RI, thus, we do not know where specifically the antibody would bind. But since the homology with mouse is quite good, the antibody might work in zebrafish.
Also, whole mount has not yet been tested with this antibody.
For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
Please let me know if you would be interested in a testing discount code.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer


I am very pleased to hear you would like to accept our offer and test ab11512 in IHC-P. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview forIHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your enquiry. The homology of human E cadherin with cow is 77%. The homology with pig is 80%. For your information, our other antibodies to E cadherin are raised against human or mouse. The homology of mouse protein with cow is 75%, with pig it is 79%. If your customer chooses a monoclonal antibody I would not expect cross reactivity. For polyclonal antibodies, there may be a small chance. Please don't hesitate to contact us if you need further assistance.

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Answer

I'm sorry to hear you are having problem with ab11512. I would like to suggest the following modifications to your protocol: -fix cells and tissue in very cold acetone (pre-cooled at -20C) for 10min, (try this on sections too) -wash 3-5x in PBS between steps -add some triton x100 to your BSA-PBS buffer that you use to dilute your antibodies in (typically 0.3%v/v), this will help the penetration of the antibody into the tissue -check your secondary antibody works with other primary antibodies -try a positive control of MDCK cells. Please let me know if this helps and do not hesitate to contact us for further advice,

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Answer

Thank you for your enquiry regarding ab11512. I'm sorry to hear you have had problems with this antibody. As you probably noticed on the datasheet this antibody has not been tested in paraffin sections, but should definitely work in frozen sections. As you have had an identical problem with another of our antibodies which should also work in your tissue I would like to refer to the same advice I gave you: -add some triton x100 to your BSA-PBS buffer that you use to dilute your antibodies in (typically 0.3%v/v), this will help the penetration of the antibody into the tissue, -incubate the primary antibody overnight at high humidity and 4C. -incubate the secondary antibody for longer, try 2 hours. Let me know if you need further help,

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