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Knockout Tested Rabbit Monoclonal E Cadherin antibody. Carrier free. Suitable for Flow Cyt, IHC-P, WB, ICC/IF, Flow Cyt (Intra), mIHC and reacts with Human samples. Cited in 7 publications.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (AB201499), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (AB201499), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (AB201499), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (AB201499), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (AB201499), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow CytIHC-PWBICC/IFFlow Cyt (Intra)mIHC
Human
Tested
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Associated Products

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Target data

Function

Cadherins are calcium-dependent cell adhesion proteins (PubMed:11976333). 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 (PubMed:11976333). Promotes organization of radial actin fiber structure and cellular response to contractile forces, via its interaction with AMOTL2 which facilitates anchoring of radial actin fibers to CDH1 junction complexes at the cell membrane (By similarity). 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. (Microbial infection) Serves as a receptor for Listeria monocytogenes; internalin A (InlA) binds to this protein and promotes uptake of the bacteria.

Alternative names

Recommended products

Knockout Tested Rabbit Monoclonal E Cadherin antibody. Carrier free. Suitable for Flow Cyt, IHC-P, WB, ICC/IF, Flow Cyt (Intra), mIHC and reacts with Human samples. Cited in 7 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EP700Y
Purification technique
Affinity purification Protein A
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. This antibody has been tested on human samples in both WB and IHC. Customer feedback (see Abreview) suggests the antibody does not perform well in IHC on mouse tissue.

Dissociation constant
2.8 x 10-11 M
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab201499 is the carrier-free version of Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772.

Mouse, Rat: We have preliminary internal testing data to indicate 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.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Our Low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

E-Cadherin sometimes called CDH1 or Cadherin-1 is a protein that plays a role in cell-to-cell adhesion. This transmembrane protein has a molecular weight of approximately 120 kDa. E-Cadherin is mainly expressed in epithelial tissues of various organs including the skin and gut. Its adhesive function is made possible by its extracellular domain that facilitates homophilic binding between cells contributing to the maintenance of tissue architecture and cellular integrity.

Biological function summary

E-Cadherin participates in establishing and maintaining adherens junctions which are vital for tissue structure. E-Cadherin operates as a core component of the cadherin-catenin complex which links the protein to the actin cytoskeleton. Through this linkage E-Cadherin plays an important role in signaling pathways that influence cellular growth and differentiation. The protein's ability to mediate intercellular connections also regulates cellular motility and supports basic aspects of cell behavior in epithelial tissues.

Pathways

E-Cadherin influences both the Wnt signaling pathway and the epithelial-to-mesenchymal transition (EMT). Within the Wnt signaling pathway E-Cadherin partners with β-catenin a significant player in transcription regulation and cell signaling. Disruption in E-Cadherin's adhesive functionality can lead to increased β-catenin availability affecting downstream transcriptional control. In the EMT process E-Cadherin loss characterizes an important step in which cells gain migratory and invasive properties typically seen during metastasis in cancer progression.

Associated diseases and disorders

E-Cadherin's role is prominent in cancer particularly in the context of gastric and breast cancers. Mutations or altered expression of E-Cadherin lead to diminished cell adhesion promoting tumorigenesis and metastatic spread. In gastric cancer for instance mutated E-Cadherin often associates with loss of epithelial function facilitating cancer cell dissemination. Additionally its loss or dysfunction may correlate with proteins such as β-catenin further impacting cancer progression and pathology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

21 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail
    Image from Roca H. et al PLoS One. 2013 Oct 4;8(10):e76773. doi: 10.1371/journal.pone.0076773. eCollection 2013.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Mesenchymal cancer cells show increased metastasis while not requiring MET for solid tumor formation.

    ZEB1 or E-cadherin staining of metastases in ICI-mice. Note the higher E-cad and lower ZEB1 expression in the metastatic cells expressing OVOL1 or ZEB1-shRNA (sh4). Scale bar represents 100 μm.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 staining E Cadherin in HT-29 (Human colorectal adenocarcinoma) cells by ICC/IF (Immunocytochemistry/Immunofluorescence).  Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at 1/500 dilution. An Alexa Fluor® 488 Goat anti-Rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 dilution. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution was used as a counterstain. DAPI was used as a nuclear counterstain. This is a confocal image showing membranous staining on HT-29 cell line.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail
    Image from Omata W. et al PLoS One. 2013 Nov 13;8(11):e81003. doi: 10.1371/journal.pone.0081003. eCollection 2013.

    Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    PMA induced cell fusion, DYSF expression, and activation of PKC in BeWo cells while 4αPMA was inactive

    Immunofluorescence analysis of BeWo cells treated with 0.25% DMSO (controls), 10 nM PMA, or 10 nM 4αPMA for 72 h. The cells were then fixed and subsequently double-labeled for detection of DYSF (red) and E-cadherin (green). Nuclei were labeled with DAPI. While there can be a low level of spontaneous fusion in control cells (in our hands this ranges from about 4 to 9%), most cells are not fused and have at their borders intact E-cadherin labeling. Moreover, DYSF labeling was not detectable in non-fused BeWo cells. However, treatment of BeWo cells with 10 nM PMA for 72 h led to increased levels of cell fusion as indicated by the breakdown of E-cadherin labeling and the expression of DYSF in fused cells. When BeWo cells were treated with 10 nM 4αPMA for 72 h there was no detectable increase in cell fusion or DYSF expression. Arrows indicate areas enlarged and placed in insets. Bar = 50 µm.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Flow Cytometry (Intracellular) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling E Cadherin with purified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at 1/30 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Immunohistochemistry of breast carcinoma staining E Cadherin with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at 1μg/ml

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Immunocytochemistry/Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial) cells labeling E Cadherin with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were then incubated with the primary antibody at a 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at a 1/1000 dilution (green). The nuclear counter stain is DAPI (blue). Counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).

    Confocal image shows membranous staining on MCF7 cell line.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Flow Cytometry (Intracellular) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Overlay histogram showing A431 (Human epidermoid carcinoma cell line) cells stained with unpurified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 (red line). The cells were fixed with 80% methanol (5 minutes) 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 (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772, 1/1000 dilution) for 30 minute at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1 μg/1x106 cells) used under the same conditions. Unlabeled 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for E Cadherin with unpurified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at a 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Formalin/PFA-fixed paraffin-embedded human colonic adenocarcinoma tissue stained for E Cadherin with unpurified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at a 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Formalin-fixed, paraffin-embedded human papillary carcinoma of thyroid gland tissue stained for E Cadherin with unpurified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at a 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Formalin-fixed, paraffin-embedded human transitional cell carcinoma of kidney tissue stained for E Cadherin with unpurified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at a 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Formalin-fixed, paraffin-embedded human lung adenocarcinoma tissue stained for E Cadherin with unpurified Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at a 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    OI-RD Scanning - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    This IHC data was generated using the same anti-E Cadherin antibody clone, EP700Y, in a different buffer formulation (cat# Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772. Green-E-Cadherin red-PI

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was as GAPDH loading control.

    Exposure time: Lane1: 3 seconds; Lane 2-5: 40 seconds.

    A375, HeLa and HT-1080 were reported as negative or express low level of E cadherin (PMID: 30393081, PMID: 16980628, PMID: 34715746), PMID: 25411788).

    All lanes: Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) at 1/1000 dilution

    Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: A375 (Human malignant melanoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 97 kDa

  • Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    False colour image of Western blot: Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 was shown to bind specifically to E Cadherin. A band was observed at 105/130 kDa in wild-type Raji cell lysates with no signal observed at this size in CDH1 knockout cell line Human CDH1 (E Cadherin) knockout A-431 cell line ab273747 (knockout cell lysate ab273781). To generate this image, wild-type and CDH1 knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) at 1/10000 dilution

    Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499) at 1/10000 dilution

    Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [EP700Y] - BSA and Azide free (Anti-E Cadherin antibody [EP700Y] - BSA and Azide free ab256580) at 1/10000 dilution

    Lane 1: Wild-type Raji cell lysate at 20 µg

    Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (Human CDH1 (E Cadherin) knockout A-431 cell line ab273747)

    Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)

    Lane 2: CDH1 knockout Raji cell lysate at 20 µg

    Lane 3: MCF7 cell lysate at 20 µg

    Lane 4: MDA-MB-231 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 105 kDa, 130 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Immunofluorescence staining of E-Cadherin using Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 in wild-type A431 cells (top panel) and CDH1 knockout A431 cells (bottom panel). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton-X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at 1 µg/mL and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a confocal section is shown.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Immunofluorescence staining of E-Cadherin using Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 in wild-type A431 cells (top panel) and CDH1 knockout A431 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton-X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at 0.2 µg/mL and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a confocal section is shown.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

  • Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    Exposure time: 3.25 seconds.

    Blocking and diluting buffer: 5% NFDM/TBST.

    Full-length E Cadherin has a molecular weight of approximately 125 kDa. Other molecular weights between 80-100 kDa could also be observed depending on cell types or cell conditions.

    PMID: 27274359, PMID: 26983597, PMID: 18478055, PMID: 22375065.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    All lanes: Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) at 1/1000 dilution

    All lanes: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

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

    Predicted band size: 97 kDa

    Observed band size: 80-125 kDa

  • Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).
    Western blot: Anti-CDH1 antibody [EP700Y] (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 was shown to bind specifically to CDH1. A band was observed at 130, 110, 80, 55, 40 kDa in wild-type A431 cell lysates with no signal observed at this size in CDH1 knockout cell line Human CDH1 (E Cadherin) knockout A-431 cell line ab273747 (knockout cell lysate ab273781). To generate this image, wild-type and CDH1 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) at 1/1000 dilution

    Lane 1: Wild-type A431 cell lysate at 20 µg

    Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)

    Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (Human CDH1 (E Cadherin) knockout A-431 cell line ab273747)

    Lane 2: CDH1 knockout A431 cell lysate at 20 µg

    Lane 3: Caco-2 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 97 kDa

  • Flow Cytometry - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499), expandable thumbnail

    Flow Cytometry - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772).

    Flow cytometry overlay histogram showing MCF7 positive cells (left) and negative MDA-MB-231 cells (right) stained with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction, followed by the antibody Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 (1x 106 in 100μl at 5 μg/ml (1/400)) for 30min on ice.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

    Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

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