Anti-E Cadherin [EP700Y] antibody (ab256580) is a rabbit monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect E Cadherin in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF, mIHC. Suitable for Human samples.
- Specificity confirmed with E Cadherin knockout cell line validation
-BSA,?sodium azide, and glycerol-free?for consistent conjugation with fluorochromes, enzymes and more
pH: 7.2 - 7.4
Constituents: PBS
Flow Cyt | mIHC | WB | IHC-P | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/3000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Select an associated product type
CDH1 encodes a protein that is a calcium-dependent cell adhesion molecule, playing a role in cell-cell adhesion, mobility, and proliferation in epithelial cells. It acts as a potent invasive suppressor and serves as a ligand for integrin alpha-E/beta-7. The protein contributes to the sorting of heterogeneous cell types due to its homophilic interactions. E-Cad/CTF2, a product of CDH1, promotes the non-amyloidogenic degradation of Abeta precursors and strongly inhibits the production of APP C99 and C83. Additionally, in the context of microbial infection, it functions as a receptor for Listeria monocytogenes, with internalin A binding to it to promote bacterial uptake. This supplementary information is collated from multiple sources and compiled automatically.
CD324, CDHE, UVO, CDH1, Cadherin-1, CAM 120/80, Epithelial cadherin, Uvomorulin, E-cadherin
Anti-E Cadherin [EP700Y] antibody (ab256580) is a rabbit monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect E Cadherin in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF, mIHC. Suitable for Human samples.
- Specificity confirmed with E Cadherin knockout cell line validation
-BSA,?sodium azide, and glycerol-free?for consistent conjugation with fluorochromes, enzymes and more
pH: 7.2 - 7.4
Constituents: PBS
ab256580 is the carrier-free version of Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772.
Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.
Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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).
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 (10μg/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 of breast carcinoma staining E Cadherin with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 at 1μg/ml.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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 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).
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).
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).
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).
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).
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).
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).
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.
This IHC data was generated using the same anti-E Cadherin antibody clone, EP700Y, in a different buffer formulation (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.
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).
Tissue Microarrays stained for Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker using Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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 (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 (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
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
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).
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).
Fluorescence multiplex immunohistochemical analysis of the human endometrium (Formalin/PFA-fixed paraffin-embedded sections).
Panel A: merged staining of anti-E Cadherin (ab256580, red; Opal™690), anti-SLC34A2 (Anti-SLC34A2 antibody [SP322] - BSA and Azide free ab238793, green; Opal™520) and anti-CD10 (Anti-CD10 antibody [EPR22865-73] ab255609, cyan; Opal™570) on human endometrium. Panel B: anti-CD10 stained on stromal cells. Panel C: anti-E Cadherin stained on glandular cells. Panel D: anti-SLC34A2 stained on apical membrane of glandular cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody.
The section was incubated in three rounds of staining: in the order of ab256580 at 1/3000 dilution (0.324 μg/ml) for 30mins, Anti-SLC34A2 antibody [SP322] - BSA and Azide free ab238793 at 1/1000 dilution (2.26 μg/ml) for 10mins and Anti-CD10 antibody [EPR22865-73] ab255609 at 1/1000 dilution (0.615 μg/ml) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain.
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
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
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
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
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
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
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.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com