Recombinant Anti-Ezrin antibody [EPR803] (ab133297)
![Western blot - Anti-Ezrin antibody [EPR803] (ab133297)](https://www.abcam.com/ps/products/133/ab133297/Images/ab133297-448916-anti-ezrin-antibody-epr803-western-blot-hela-lysates.jpg "Western blot - Anti-Ezrin antibody [EPR803] (ab133297)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR803] to Ezrin
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Ezrin antibody [EPR803]
See all Ezrin primary antibodies -
Description
Rabbit monoclonal [EPR803] to Ezrin -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Ezrin aa 350-450. The exact sequence is proprietary.
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Positive control
- WB: HAP1, HeLa, HCT116, 293T, A549 and SH-SY5Y cell lysates. IHC-P: Human kidney tissue and Human transitional cell carcinoma tissue.
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General notes
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR803 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133297 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/10000 - 1/50000. Detects a band of approximately 81 kDa (predicted molecular weight: 69 kDa).
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| IHC-P |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| Notes |
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WB
1/10000 - 1/50000. Detects a band of approximately 81 kDa (predicted molecular weight: 69 kDa). |
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IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
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. -
Tissue specificity
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. -
Sequence similarities
Contains 1 FERM domain. -
Developmental stage
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. -
Post-translational
modificationsPhosphorylated by tyrosine-protein kinases. -
Cellular localization
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
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Database links
- Entrez Gene: 7430 Human
- Omim: 123900 Human
- SwissProt: P15311 Human
- Unigene: 487027 Human
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Alternative names
- Villin 2 ezrin antibody
- CVIL antibody
- CVL antibody
see all
Images
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Western blot - Anti-Ezrin antibody [EPR803] (ab133297)All lanes : Anti-Ezrin antibody [EPR803] (ab133297) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EZR knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab133297 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133297 was shown to react with Ezrin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264912 (knockout cell lysate ab256908) was used. Wild-type HeLa and EZR knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133297 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)](https://www.abcam.com/ps/products/133/ab133297/Images/ab133297-146781-anti-ezrin-antibody-epr803-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Ezrin with ab133297 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Western blot - Anti-Ezrin antibody [EPR803] (ab133297)](https://www.abcam.com/ps/products/133/ab133297/Images/ab133297-296521-anti-ezrin-antibody-epr803-western-blot.jpg)
Western blot - Anti-Ezrin antibody [EPR803] (ab133297)All lanes : Anti-Ezrin antibody [EPR803] (ab133297) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Ezrin knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HCT116 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 69 kDaLanes 1 - 4: Merged signal (red and green). Green - ab133297 observed at 75 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab133297 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. Ab133297 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)](https://www.abcam.com/ps/products/133/ab133297/Images/ab133297-146780-anti-ezrin-antibody-epr803-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma tissue labelling Ezrin with ab133297 at 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Western blot - Anti-Ezrin antibody [EPR803] (ab133297)](https://www.abcam.com/ps/products/133/ab133297/Images/ab133297-146779-anti-ezrin-antibody-epr803-western-blot.jpg)
Western blot - Anti-Ezrin antibody [EPR803] (ab133297)All lanes : Anti-Ezrin antibody [EPR803] (ab133297) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : A549 cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-rabbit HRP at 1/2000 dilution
Predicted band size: 69 kDa -
![Anti-Ezrin antibody [EPR803] (ab133297)](https://www.abcam.com/ps/products/133/ab133297/Images/ab133297-7-benefits-of-recombinant-antibodies.png)
Anti-Ezrin antibody [EPR803] (ab133297)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab133297 has not yet been referenced specifically in any publications.