Name: Anti-E Cadherin antibody [4A2] - BSA and Azide free
SKU: ab233766
Rating: 5 (1 reviews)

Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766) is a mouse monoclonal antibody provided in a PBS only buffer for easy conjugation detecting E Cadherin in Western Blot, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat.

- KO validated for confirmed specificity

- BSA, sodium azide, and glycerol-free for easy conjugation

Images

Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [4A2] - BSA and Azide free (AB233766), expandable thumbnail

Publications

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: 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

	IHC-P	WB	ICC/IF
Human	Tested	Tested	Tested
Mouse	Tested	Tested	Expected
Rat	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1 µg/mL	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1 µg/mL	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1 µg/mL	Notes Abcam recommends using a 5% milk blocking solution for this antibody.
Species Rat	Dilution info 1 µg/mL	Notes Abcam recommends using a 5% milk blocking solution for this antibody.
Species Human	Dilution info 1 µg/mL	Notes Abcam recommends using a 5% milk blocking solution for this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1 µg/mL	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

Select an associated product type

1 product for Alternative Version

7 products for Alternative Product

Target data

See full target information CDH1

Function

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.

Alternative names

CD324, CDHE, UVO, CDH1, Cadherin-1, CAM 120/80, Epithelial cadherin, Uvomorulin, E-cadherin

Key facts

Isotype: IgG1
Host species: Mouse
Storage buffer: Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: 4A2
Purification technique: Affinity purification
Light chain type: kappa
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Aliquoting information: Upon delivery aliquot
Storage information: Do Not Freeze

Notes

What is this antibody validated in?

Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766) is a mouse monoclonal antibody and is validated for use in Western Blot (WB), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.

What is the molecular weight of E Cadherin?

Anti-E Cadherin [4A2] - BSA and Azide free (ab233766) specifically detects a band for E Cadherin (UniProt: P12830) at a molecular weight of 97kDa.

Specificity confirmed

The specificity of Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766) has been confirmed by Western blot testing in CDH1 Knockout A431 cells.

Other related products

We have a range of other formats of antibody clone [4A2] also available for your convenience:
ab231303, Carrier free - ab233766

Want a custom formulation?
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com

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.

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.

8 product images

Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
Immunofluorescence staining of E-Cadherin using Anti-E Cadherin antibody [4A2] ab231303 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 [4A2] ab231303 at 1.0 µg/mL and Anti-beta Tubulin antibody - Loading Control ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (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 and sodium azide (Anti-E Cadherin antibody [4A2] ab231303).
Western blot - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
This data was developed using the same antibody clone in a different buffer formulation (Anti-E Cadherin antibody [4A2] ab231303).
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before Anti-E Cadherin antibody [4A2] ab231303 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/10000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-E Cadherin antibody [4A2] (Anti-E Cadherin antibody [4A2] ab231303)
Lane 1: MCF7 whole cell lysate at 20 µg
Lane 2: Mouse colon tissue lysate at 20 µg
Lane 3: Rat colon tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 97 kDa
Observed band size: 105 kDa
Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
Anti-E Cadherin antibody [4A2] ab231303 staining E-Cadherin in MCF7 cells. The cells were fixed with 4% PFA (10min), permeabilized with 0.1% PBS-Tween 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 overnight at +4°C with Anti-E Cadherin antibody [4A2] ab231303 at 1μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This data was developed using the same antibody clone (Anti-E Cadherin antibody [4A2] ab231303) in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
IHC image of E Cadherin staining in a section of formalin-fixed paraffin-embedded normal human colon carcinoma* performed on a Leica BOND^TM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Anti-E Cadherin antibody [4A2] ab231303, 1 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-E Cadherin antibody [4A2] ab231303).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
This data was developed using ab233766, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of formalin fixed paraffin embedded human colon carcinoma labelling E cadherin with Anti-E Cadherin antibody [4A2] ab231303 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
Anti-E Cadherin antibody [4A2] ab231303 anti-E Cadherin antibody [4A2] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).
Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
Immunofluorescence staining of E-Cadherin using Anti-E Cadherin antibody [4A2] ab231303 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 [4A2] ab231303 at 1.0 µg/mL and Anti-beta Tubulin antibody - Loading Control ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084) (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 and sodium azide (Anti-E Cadherin antibody [4A2] ab231303).
Western blot - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
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 [4A2] ab231303).
Western blot: Anti-CDH1 antibody [4A2] (Anti-E Cadherin antibody [4A2] ab231303) staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-E Cadherin antibody [4A2] ab231303 was shown to bind specifically to CDH1. A band was observed at 130, 110, 55 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 5 % 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-E Cadherin antibody [4A2] (Anti-E Cadherin antibody [4A2] ab231303) at 1 µg/mL
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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 97 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [4A2] - BSA and Azide free (ab233766)
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (Anti-E Cadherin antibody [4A2] ab231303).

IHC image of E-Cadherin staining in a section of formalin-fixed paraffin-embedded normal mouse large intestine performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with Anti-E Cadherin antibody [4A2] ab231303, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.