Anti-E Cadherin 抗体 [EP700Y] - Intercellular Junction Marker (ab40772)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP700Y] to E Cadherin - Intercellular Junction Marker
- Suitable for: ICC/IF, IHC-P, Flow Cyt, WB
- Reacts with: Human
製品の概要
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製品名
Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker
E Cadherin 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP700Y] to E Cadherin - Intercellular Junction Marker -
由来種
Rabbit -
特異性
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. -
アプリケーション
適用あり: ICC/IF, IHC-P, Flow Cyt, WBmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human E Cadherin aa 600-700. The exact sequence is proprietary.
Database link: P12830 -
ポジティブ・コントロール
- WB: Positive controls: MCF-7, HT-29, PC-3 whole cell lysate. Negative Control: MDA-MB-231 whole cell lysate IHC-P: Human breast carcinoma, lung adenocarcinoma and colonic adenocarcinoma tissue. Human papillary carcinoma of thyroid gland and transitional cell carcinoma of kidney tissue. ICC/IF: MCF7 and HT-29 cells. Flow Cytometry: A431 and MCF7 cells.
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特記事項
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
解離定数(KD 値)
KD = 2.80 x 10 -11 M Learn more about KD -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP700Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Alexa Fluor® 488 Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab185013)
- Alexa Fluor® 647 Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab194982)
- Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (ab201499)
- Alexa Fluor® 555 Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab206878)
- Alexa Fluor® 594 Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab206880)
- PE Anti-E Cadherin antibody [EP700Y] (ab224959)
- APC Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab224960)
- Anti-E Cadherin antibody [4A2] (ab231303)
- Anti-E Cadherin antibody [EP700Y] - BSA and Azide free (ab256580)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
Our Abpromise guarantee covers the use of ab40772 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | 1/500. Permeabilisation is unnecessary as the immunogen is in an extracellular domain. |
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IHC-P | 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
Flow Cyt | 1/30. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For purified format use at 1/1000. |
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WB | 1/10000 - 1/50000. Detects a band of approximately 80-120 kDa (predicted molecular weight: 97 kDa). |
ターゲット情報
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機能
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. -
組織特異性
Non-neural epithelial tissues. -
関連疾患
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. -
配列類似性
Contains 5 cadherin domains. -
翻訳後修飾
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. -
細胞内局在
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
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参照データベース
- Entrez Gene: 999 Human
- Omim: 192090 Human
- SwissProt: P12830 Human
- Unigene: 461086 Human
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別名
- Arc 1 antibody
- CADH1_HUMAN antibody
- Cadherin 1 antibody
see all
画像
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All lanes : Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/10000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell). Whole cell lysates
Lane 2 : HT-29 (Human colorectal adenocarcinoma epithelial cell). Whole cell lysates
Lane 3 : PC-3 (Human prostate adenocarcinoma epithelial cell) Whole cell lysates
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) Whole cell lysates (negative control)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Performed under reducing conditions.
Predicted band size: 97 kDa
Exposure time: 23 secondsBlocking and diluting buffer: 5% NFDM/TBST
Multi-bands can refer to PMID: 11212238; PMID: 14695147 and PMID: 22659456
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Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)Omata, W. et al PLoS One. 2013 Nov 13;8(11):e81003. doi: 10.1371/journal.pone.0081003. eCollection 2013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)Roca, H. et al PLoS One. 2013 Oct 4;8(10):e76773. doi: 10.1371/journal.pone.0076773. eCollection 2013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
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.
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Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
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 (ab150077) was used as the secondary antibody at 1/1000 dilution. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (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.
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Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling E Cadherin with purified ab40772 at 1:30 dilution (10 µg/ml) (red). 106 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).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
Fluorescent immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab40772. Green-E-Cadherin red-PI.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Overlay histogram showing A431 (Human epidermoid carcinoma cell line) cells stained with unpurified ab40772 (red line). 106 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 (ab40772, 1/1000 dilution) for 30 minute at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
Formalin/PFA-fixed paraffin-embedded human colonic adenocarcinoma tissue stained for E Cadherin with unpurified ab40772 at a 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Lane 1 : Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/5000 dilution
Lane 2 : Anti-E Cadherin antibody [EPR699] (ab133597) at 1/2000 dilution
Lane 3 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
All lanes : PC-3 (Human prostate adenocarcinoma epithelial cell) Whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 97 kDaExposure time: 3 minutes for ab40772 and ab133597, 32 seconds for GAPDH.
Blocking and diluting buffer: 5% NFDM/TBST
Multi-bands can refer to PMID: 11212238; PMID: 14695147 and PMID: 22659456
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Immunocytochemistry/ Immunofluorescence - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
Immunocytochemistry/Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial) cells labeling E Cadherin with 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) (ab150077) secondary antibody at a 1/1000 dilution (green). The nuclear counter stain is DAPI (blue). Counterstained with 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
Formalin-fixed, paraffin-embedded human lung adenocarcinoma tissue stained for E Cadherin with unpurified ab40772 at a 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Lane 1 : Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/5000 dilution
Lane 2 : Anti-E Cadherin antibody [EPR699] (ab133597) at 1/2000 dilution
Lane 3 : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
All lanes : HT-29 (Human colorectal adenocarcinoma epithelial cell). Whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 97 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?Exposure time: 1 second for ab40772, 3 minutes for ab133597, 32 seconds for GAPDH
Blocking and diluting buffer: 5% NFDM/TBST
Multi-bands can refer to PMID: 11212238; PMID: 14695147 and PMID: 22659456
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for E Cadherin with unpurified ab40772 at a 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772) at 1/200000 dilution (Unpurified) + MCF7 (Human breast adenocarcinoma ) whole cell lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) at 1/1000 dilution
Predicted band size: 97 kDa
Observed band size: 100,120,80,97 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking and diluting buffer and concentration 5% NFDM/TBST.
The full-length of E-cadherin is 120 kDa. The other bands are due to proteolytic cleavages in different Cadherin domains. (Ref: PMID: 14695147)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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 ab40772 at a 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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 ab40772 at a 1/500 dilution in immunohistochemical analysis.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (ab40772)
Immunohistochemistry of breast carcinoma staining E Cadherin with ab40772 at 1μg/ml
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Produced using unpurified ab40772
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
参考文献 (262)
ab40772 は 262 報の論文で使用されています。
- Zhao W et al. Splicing factor derived circular RNA circUHRF1 accelerates oral squamous cell carcinoma tumorigenesis via feedback loop. Cell Death Differ 27:919-933 (2020). PubMed: 31570856
- Zhou S et al. Endoplasmic reticulum stress regulates epithelial-mesenchymal transition in human lens epithelial cells. Mol Med Rep 21:173-180 (2020). PubMed: 31746423
- Ke B et al. Sonic Hedgehog/Gli1 Signaling Pathway Regulates Cell Migration and Invasion via Induction of Epithelial-to-mesenchymal Transition in Gastric Cancer. J Cancer 11:3932-3943 (2020). PubMed: 32328197
- Mei B et al. The regulatory mechanism and biological significance of the Snail-miR590-VEGFR-NRP1 axis in the angiogenesis, growth and metastasis of gastric cancer. Cell Death Dis 11:241 (2020). PubMed: 32303680
- Bian S miR-4319 inhibited the development of thyroid cancer by modulating FUS-stabilized SMURF1. J Cell Biochem 121:174-182 (2020). PubMed: 31148243