Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab12221 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 1/100.
WB Use a concentration of 5 - 10 µg/ml. Use under non reducing condition. Predicted molecular weight: 99.9 kDa.
IHC-Fr Use a concentration of 5 - 10 µg/ml.
IHC-P Use a concentration of 5 - 10 µg/ml.

Target

Anti-N Cadherin antibody images

  • Paraffin-embedded sections of 15-day mouse embryo. Antigen retrieval: microwave treatment, 3 minutes x 10 times, 10 mM citrate buffer. Primary antibody: 10 ug/ml ab12221, 1 hour, room temperature. Secondary antibody: Envision plus Colorimetric detection: DAB
  • All lanes : Anti-N Cadherin antibody (ab12221) at 1/100 dilution

    Lane 1 : Whole cell lysate prepared from human glioblastoma cell line U373MG
    Lane 2 : Whole cell lysate prepared from human glioblastoma cell line U251

    Lysates/proteins at 100 µg per lane.

    Secondary
    HRP monoclonal anti-rabbit immunoglobulins at 1/7500 dilution
    Developed using the ECL technique

    Predicted band size : 99.9 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 minutes

    Image courtesy of an anonymous Abreview.

    Distinct bands were found in all tested cell lines with no unspecific bands.

    See Abreview

  • ab12221, at 1/100, staining N Cadherin in mouse differentiated embryonic stem cells by immunocytochemistry / immunofluorescence. Cells were methanol/acetone (1/1) fixed and permeabilized in 0.1% triton X prior to blocking in 1% serum and 0.1% BSA for 30 minutes at 21°C. Alexa fluor® 488 goat polyclonal to rabbit Ig, diluted 1/200, was used as the secondary antibody.

    See Abreview

  •  ab12221 at 1/200 dilution staining rat heart tissue sections by IHC-P. The tissue was fixed with Histochoice and paraffin sectioned prior to staining. An Alexa-Fluor conjugated donkey anti-rabbit antibody was used as the secondary.

    The image depicts stained N-Cadherin (green) and counterstained nuclei (DAPI-blue) overlay (left panel). The DIC (phase) image of the same tissue, superimposed with fluorescence overlay (right panel) confirms the localization of the N-Cadherin at intercalated discs region. Note, the phase contrast image shows typical uninucleated, cylindrical morphology of cardiac myocytes; that are often branched and joined to each other end-to-end (intercalated discs) to form interlacing network.

    See Abreview

  • All lanes : Anti-N Cadherin antibody (ab12221) at 1/100 dilution

    Lane 1 : MDA-MB-231 cells (Negative Control)
    Lane 2 : U2OS cells (Positive Control)

    Lysates/proteins at 200000 cells per lane.

    Secondary
    Goat anti-rabbit HRP at 1/2000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 99.9 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 75 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Immunohistochemical analysis of Human lung tissue, with idiopathic pulmonary fibrosis, staining N Cadherin with ab12221.

    Tissue was fixed in aceteone/methanol (1:1) and incubated with primary antibody overnight at 4°C. An Oregon Green 488-conjugated anti-rabbit IgG was used as the secondary antibody.

References for Anti-N Cadherin antibody (ab12221)

This product has been referenced in:
  • Hwang PY  et al. N-cadherin is Key to Expression of the Nucleus Pulposus Cell Phenotype under Selective Substrate Culture Conditions. Sci Rep 6:28038 (2016). ICC/IF . Read more (PubMed: 27292569) »
  • Verrill C  et al. Altered expression of epithelial-to-mesenchymal transition proteins in extraprostatic prostate cancer. Oncotarget 7:1107-19 (2016). Read more (PubMed: 26701730) »

See all 33 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Horse Tissue sections (aglandular stomach, epithelium)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization No
Specification aglandular stomach, epithelium
Blocking step Milk as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 222°C
Fixative methacarn, PFA
Username

Dr. F Hernandez-Blazquez

Verified customer

Submitted Jun 13 2016

Thank you for contacting us.

I am pleased to provide the following immunogen information for ab98952 N Cadherin antibody [5D5] :

Immunogen sequence: ##########

I am sorry to confirm that permeabilization was required wh...

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Thank you for contacting Abcam.
Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.
We are happy to reassure our customers that all of our products are covered by our Abpromise, which gu...

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DISCOUNT CODE: ABTRIAL-EBAMG
Expiration date: 2013-03-19
Value: CAD388.00
I am very pleased to hear you would like to accept our offer and test ab8211 in zebrafish. This code will give you CAD388.00 off your next order before the expiratio...

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Thank you very much for contacting us.
Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.
We are happy to reassure our customers that all of our products are covered by our Abpromise, whi...

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I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Cardiomyocytes)
Specification Cardiomyocytes
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Mar 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 20 µg
Specification HEK293
Gel Running Conditions Non-reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Feb 01 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (NIH3T3 cells)
Loading amount 5 µg
Specification NIH3T3 cells
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Nov 03 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (LoVo cells)
Loading amount 5 µg
Specification LoVo cells
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 21 2011

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