Overview

  • Product name
    Anti-E Cadherin antibody [HECD-1]
    See all E Cadherin primary antibodies
  • Description
    Mouse monoclonal [HECD-1] to E Cadherin
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, ICC, IP, IHC-P, IHC-Fr, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Does not react with: Mouse
  • Immunogen

    Breast tumor cell line MCF-7

  • Positive control
    • WB: MDA-MB-468 and LNCaP whole cell lysates; MCF7 cell lysate. ICC/IF: Human renal tubular epithelial cells; Human nasal epithelial cells; NSCLC cells; Human renal proximal tubular cells; LNCaP cells. IHC-P: Breast cancer tissue; Human stomach tissue. Flow Cyt: MCF7 and A431 cells. IHC-Fr: Human skin tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab1416 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 Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/50 - 1/100.
ICC/IF Use at an assay dependent concentration. PubMed: 19244118
WB 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

  • Anti-E Cadherin antibody [HECD-1] (ab1416) at 1/50 dilution + LNCaP (human prostate cancer cell line) whole cell lyate

    See Abreview

  • All lanes : Anti-E Cadherin antibody [HECD-1] (ab1416) at 1/1000 dilution

    All lanes : MDA-MB-468 cells whole cell lysate

    Lysates/proteins at 80 µg/ml per lane.

    Secondary
    All lanes : HRP-conjugated polyclonal donkey anti-mouse IgG

    Performed under reducing conditions.

    Observed band size: 110 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds

    See Abreview

  • ab1416 staining E cadherin in Human renal tubular epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 1% BSA for 1 hour at 20°C. Samples were incubated with primary antibody (1/200 in PBS + 0.01% BSA) for 24 hours at 4°C. An Alexa Fluor® 555-conjugated Donkey anti-mouse IgG polyclonal was used as the secondary antibody (1/500).

    See Abreview

  • Breast cancer stained with E Cadherin mouse antibody ab1416

  • Lanes 1-2 : Anti-E Cadherin antibody [HECD-1] (ab1416) at 1/50 dilution
    Lanes 3-4 : Anti-GAPDH at 1 µg/ml

    Lanes 1 & 3 : MDA-MB-231 cell lysate
    Lanes 2 & 4 : MCF-7 cell lysate


    Blocking buffer: 3% milk

  • Overlay histogram showing MCF7 cells stained with ab1416 (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 (ab1416, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 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. Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • ab1416 staining E Cadherin in Human nasal epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed paraformaldehyde and permeabilized with Triton X-100 0.1%. Samples were incubated with primary antibody (1/50) for 18 hours at 4°C. A Cy2®-conjugated Horse anti-mouse monoclonal(1/100) was used as the secondary antibody.

    See Abreview

  • Flow cytometric analysis of A431 (human epidermoid carcinoma cell line) cell line labeling E Cadherin with ab1416 at 1/100 dilution (2) compared with an isotype control (1).

    See Abreview

  • ab1416 staining E Cadherin in human normal skin tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with acetone. Samples were blocked with 10% serum for 30 minutes at 21°C followed by incubation with the primary antibody at a 1/50 dilution for 30 minutes at 21°C. An undiluted HRP-conjugated goat anti-mouse IgG polyclonal was used as secondary antibody.

    See Abreview

  • ab1416 staining E Cadherin in the Human NSCLC cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.3% Triton in PBS and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in 0.3% Triton, 5% Goat Serum in PBS) for 1 hour. An Alexa Fluor® 555-conjugated Goat polyclonal was used as the secondary antibody (1/500).

    See Abreview

  • ab1416 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 diluted 1/100 and incubated with sample for 1 hour at 23°C. A HRP conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

    See Abreview

  • ab1416 staining E Cadherin in Human Renal Proximal Tubular cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton ×100 and blocking with 1% BSA was done for 30 minutes at 210C. Samples were incubated with primary antibody (1/50: in 0.1 % BSA in PBS) for 24 hours at 4°C. An Alexa Fluor®488-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/1000 dilution.

    See Abreview

  • Formaldehyde-fixed, 0.2% Triton X-100 permeabilized LNCaP (human prostate cancer cell line) cells stained for E Cadherin (green) using ab1416 at 1/50 dilution in ICC/IF. Blue: Hoechst 33342 staining.

    See Abreview

  • Immunohistochemistry of breast carcinoma staining E Cadherin with ab1416 at 5μg/ml

References

This product has been referenced in:
  • Rhys AD  et al. Loss of E-cadherin provides tolerance to centrosome amplification in epithelial cancer cells. J Cell Biol 217:195-209 (2018). WB, IF ; Human . Read more (PubMed: 29133484) »
  • Kaplan N  et al. EphA2/Ephrin-A1 Mediate Corneal Epithelial Cell Compartmentalization via ADAM10 Regulation of EGFR Signaling. Invest Ophthalmol Vis Sci 59:393-406 (2018). Read more (PubMed: 29351356) »
See all 166 Publications for this product

Customer reviews and Q&As

1-10 of 22 Q&A

Answer

Thank you for sending the image. It may well be that the vial you have received was a bad vial which is why the antibody is not detecting the target although it is very strictly qc tested. If you are interested in trying the same antibody I can send you a free of charge replacement vial. A credit note or refund can also be provided if you are happy with the result of ab53033. I would also like to encourage you to submit an Abreview for ab53033; you will be awarded Abpoints that can be exchanged with Amazon vouchers. Let me know how you would like to proceed.

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Answer

Muchas gracias por enviarnos el cuestionario relleno con la información del anticuerpo.

Siento que el anticuerpo no haya dado los resultados esperados en ninguna de las técnicas utilizadas, y espero poder ayudar a su optimización. En caso contrario, si estuviera el anticuerpo aun bajo garantía, le enviaríamos un reemplazo del mismo o un reembolso del importe invertido.

Le pediría por favor que nos mandara en número de pedido con el que se adquirió el anticuerpo, y si fuera posible una o varias imágenes de los resultados obtenidos con él. De esta manera nos ayudara mejor a comprender los resultados inesperados.

Atendiendo al protocolo enviado, quisiera hacer las siguientes sugerencias:

- Nosotros solemos recomendar utilizar RIPA como buffer de lisis, por tener un mayor poder desnaturalizante que otros buffers.

-No se menciona el agente de bloqueo usado para bloquear uniones inespecíficas de la membrana. Para este anticuerpo se ha comprobado que el uso de leche al 5% durante 1 hora da buenos resultados.

- Si tienen ocasión usar tejido de carcinoma de mama como control positivo.

Espero que estas sugerencias mejoren el rendimiento del anticuerpo. De no ser así, no dude en contactarme de nuevo.

Que pase un buen día.

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Answer

Thank you for contacting us.

A clone number is given to an antibody produced by a single clone of hybridoma cells. Each hybridoma cell clone produces one single pure homogeneous type of antibody. The term "monoclonal" pertains to a single clone of cells, a single cell and the progeny of that cell. Monoclonal antibodies can be made in large amounts in the laboratory.

Each clone number represents a specific cell line cloned from ascites that was used to manufacture the antibody. Since antibodies are produced by more than one host, each cloned cell line receives a unique clone number. The clone number is not synonymous with the lot number which is in relation with the creation of the vial. There is little to no difference amongst the quality of the clones therefore, any of these 3 antibodies would work for your purposes in WB and ICC in Human.

I would however recommend ab1416 out of the three antibodies since we have had lots of postive customer reviews when using this antibody in ICC and WB.

The prices of the antibodies are as follows:

ab777: €‘370/£227 for 250 ul at approx 1-3 mg/ml

ab1416: €‘410/£252 for 500 ul at 0.05-0.066mg/ml

ab8993: €‘465/£283 for 100 ug at 1mg/ml


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Question
Answer

Thank you very much for taking the time to complete our questionnaire and sending the image.

I am happy to give some suggestions that may help optimize the results from this antibody:

- In order to obtain a clearer blot ensuring denaturing / reducing conditions is crucial. For that, use RIPA buffer as it has a high denaturing level than the buffer used.

-Run a no primary control to make sure the unspecific bands do not correspond to the secondary, and use more diluted secondary (1/5000).

- The use of a positive control is a good indicator of the antibody’s performance. For this particular case, Breast Cancer lysates are recommended as positive control.

All of our products are covered by our Abpromise guarantee; which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase. Could you please provide the purchase order number?

I hope these tips help, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again and I’ll be more than happy to help you further.

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Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with our products.

It is very difficult for us to understand the problem without further details. In order to monitor the product’s quality, and better understand the problem, I would appreciate if you could please send us the attached questionnaire completed. It only takes about 5 minutes to fill out; any images would also be very helpful.

All of our products are covered by our Abpromise® guarantee, which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

To read about our Abpromise policy: https://www.abcam.com/index.html?pageconfig=abpromise

I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us.

I have found some references which support that the pan Cytokeratin and E Cadherin would be expressed by epidermal keratinocytes:

http://jmm.sgmjournals.org/content/56/7/971.full

http://www.univet.es/_bibliography/development_characterization_of_a_canine_skin.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC275417/

http://jcb.rupress.org/content/117/2/415

If your customer is interested in using an antibody to differentiate between these cells and other epithelial cells I would suggest that they do a review of the literature to see which markers have been used in their field of research for these purposes. If they have a particular target they are interested in please do let me know and I will see if I can suggest a suitable antibody.

I hope this information has been of help. If you require any further information, please do not hesitate to contact us again.

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Answer

Thank you very much for your emails as well as all this information.xx

Both antibodies ab40772 and ab11512 are indeed a good choice for immunofluorescence. If however you need to stain dog cells, I would recommend the ab11512 as the ab40772 has not yet been tested on dog cells to our knowledge. It could therefore be that the ab40772 would not work on these cells.
https://www.abcam.com/index.html?datasheet=40772 (or use the following: https://www.abcam.com/index.html?datasheet=40772).
https://www.abcam.com/index.html?datasheet=11512 (or use the following: https://www.abcam.com/index.html?datasheet=11512).

Please let me know whether you have any questions and confirm which antibody you would like to receive.

Thank you for your cooperation. I am looking forward to hear back from you.

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Answer

Thank you for your message.

I am sorry to hear the antibody ab1416 has stopped workingafter only one month,stored at 4ºC as recommended on the datasheet andI would be pleased to arrange in compensation either:
- a replacement with a different lot number of ab1416,
-a replacement with an alternative anti-E cadherinfrom this list : https://www.abcam.com/index.html?t=7817&pt=1
- a credit note.

Could please indicate how you would like to proceed?

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Answer

Thank you for your inquiry and for stopping by our booth at AACR.

The immunogen that was used was MCF-7 breast cancer cell line. We know that the antibody binds to the extracellular domain of E cadherin, but the actual epitope sequencehas not been mapped. The extracellular domain is from a.a.155 - 709, according to Swiss Prot.

http://www.uniprot.org/uniprot/P12830

I am sorry that this information could not be more helpful, but please contact us with any other questions.

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1-10 of 22 Q&A

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