Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP886Y] to Ezrin - Plasma Membrane Marker
- Suitable for: ICC/IF, WB, IP, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
产品名称Anti-Ezrin抗体[EP886Y] - Plasma膜Marker
参阅全部 Ezrin 一抗
描述兔单克隆抗体[EP886Y] to Ezrin - Plasma膜Marker
经测试应用适用于: ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
Synthetic peptide within Human Ezrin aa 450-550 (C terminal). The exact sequence is proprietary.
- WB: Wild-type HAP1 whole cell lysate; HeLa and HCT116 whole cell lysates. IHC-P: Human Colon, breast and bladder carcinoma. ICC/IF: HeLa cells Flow Cyt: HeLa cells IP: HeLa cells
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
纯度Protein A purified
Our Abpromise guarantee covers the use of ab40839 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
For upurified use at 1/50 - 1/100
|WB||1/5000 - 1/50000. Detects a band of approximately 72 kDa (predicted molecular weight: 69 kDa).|
For upurified use at 1/50
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
For upurified use at 1/1000
|IHC-P||1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
功能Probably involved in connections of major cytoskeletal structures to the plasma membrane. In epithelial cells, required for the formation of microvilli and membrane ruffles on the apical pole. Along with PLEKHG6, required for normal macropinocytosis.
组织特异性Expressed in cerebral cortex, basal ganglia, hippocampus, hypophysis, and optic nerve. Weakly expressed in brain stem and diencephalon. Stronger expression was detected in gray matter of frontal lobe compared to white matter (at protein level). Component of the microvilli of intestinal epithelial cells. Preferentially expressed in astrocytes of hippocampus, frontal cortex, thalamus, parahippocampal cortex, amygdala, insula, and corpus callosum. Not detected in neurons in most tissues studied.
序列相似性Contains 1 FERM domain.
发展阶段Very strong staining is detected in the Purkinje cell layer and in part of the molecular layer of the infant brain compared to adult brain.
翻译后修饰Phosphorylated by tyrosine-protein kinases.
细胞定位Apical cell membrane. Cell projection. Cell projection > microvillus membrane. Cell projection > ruffle membrane. Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. Localization to the apical membrane of parietal cells depends on the interaction with MPP5. Localizes to cell extensions and peripheral processes of astrocytes (By similarity). Microvillar peripheral membrane protein.
- Information by UniProt
- Villin 2 ezrin antibody
- CVIL antibody
- CVL antibody
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Ezrin knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HCT116 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab40839 observed at 75 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab40839 was shown to specifically react with Ezrin in wild-type HAP1 cells as signal was lost in Ezrin knockout cells. Wild-type and Ezrin knockout samples were subjected to SDS-PAGE. Ab40839 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling Ezrin with Purified ab40839 at 1:250 dilution (3.28 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ezrin with Purified ab40839 at 1:500 dilution (1.6 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
All lanes : Anti-Ezrin antibody [EP886Y] - Plasma Membrane Marker (ab40839) at 1/20000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 69 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling Ezrin with Purified ab40839 at 1:250 dilution (3.28 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ezrin with Purified ab40839 at 1:800 dilution (1 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
ab40839 (purified) at 1:40 dilution (2µg) immunoprecipitating Ezrin in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab40839 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40839 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
Overlay histogram showing SH-SY5Y cells stained with upurified ab40839 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40839, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled 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 antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ICC/IF image of unpurified ab40839 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab40839 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Anti-Ezrin antibody [EP886Y] - Plasma Membrane Marker (ab40839) at 1/50000 dilution (Unpurified) + Hela cell lysate at 10 µg
Predicted band size: 69 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?
Unpurified ab40839 at a 1:100 dilution staining Ezrin in human colon carcinoma tissue.
Fluorescent immunohistochemical analysis of paraffin-embedded human kidney carcinoma tissue using unpurified ab40839. Green-Ezrin red-PI
Unpurified ab40839 showing positive staining in Breast carcinoma tissue.
Unpurified ab40839 showing positive staining in Prostatic carcinoma tissue.
Unpurified ab40839 showing positive staining in Hepatocellular carcinoma tissue.
Unpurified ab40839 showing positive staining in Normal kidney tissue.
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.
ab40839 被引用在 6 文献中.
- Schumacher D et al. Heterogeneous pathway activation and drug response modelled in colorectal-tumor-derived 3D cultures. PLoS Genet 15:e1008076 (2019). PubMed: 30925167
- Zhang Y & Wang G MicroRNA-183 inhibits A375 human melanoma cell migration and invasion by targeting Ezrin and MMP-9. Oncol Lett 17:548-554 (2019). PubMed: 30655800
- Yan H et al. Upregulation of miR-183-5p is responsible for the promotion of apoptosis and inhibition of the epithelial-mesenchymal transition, proliferation, invasion and migration of human endometrial cancer cells by downregulating Ezrin. Int J Mol Med 42:2469-2480 (2018). PubMed: 30226564
- Ma L & Jiang T Clinical implications of Ezrin and CD44 co-expression in breast cancer. Oncol Rep 30:1899-905 (2013). PubMed: 23900701
- Grzendowski M et al. Differential proteome analysis of human gliomas stratified for loss of heterozygosity on chromosomal arms 1p and 19q. Neuro Oncol 12:243-56 (2010). PubMed: 20167812
- Wang G et al. MicroRNA-183 regulates Ezrin expression in lung cancer cells. FEBS Lett 582:3663-8 (2008). PubMed: 18840437