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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Binding Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Ezrin antibody [EPR803] (ab133297)

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Western blot - Anti-Ezrin antibody [EPR803] (ab133297)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)
  • Western blot - Anti-Ezrin antibody [EPR803] (ab133297)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)
  • Western blot - Anti-Ezrin antibody [EPR803] (ab133297)
  • 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

You may also be interested in

Protein
Product image
Recombinant Human Ezrin protein (ab132942)
Knockout
Product image
Human EZR (Ezrin) knockout HeLa cell lysate (ab256908)

View more associated products

Overview

  • 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.

  • Positive control

    • WB: HAP1, HeLa, HCT116, 293T, A549 and SH-SY5Y cell lysates. IHC-P: Human kidney tissue and Human transitional cell carcinoma tissue.
  • General notes

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

Properties

  • 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

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR803
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Binding Proteins
    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Cancer
    • Signal transduction
    • Other
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Other

Associated products

  • Alternative Versions

    • Anti-Ezrin antibody [EPR803] - BSA and Azide free (ab248452)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human EZR (Ezrin) knockout HeLa cell line (ab264912)
  • KO cell lysates

    • Human EZR (Ezrin) knockout HeLa cell lysate (ab256908)
  • Positive Controls

    • HeLa membrane extract lysate (ab29547)
  • Recombinant Protein

    • Recombinant Human Ezrin protein (ab132942)

Applications

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).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    • 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
      modifications

      Phosphorylated 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.
    • Target information above from: UniProt accession P15311 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 7430 Human
      • Omim: 123900 Human
      • SwissProt: P15311 Human
      • Unigene: 487027 Human
      • Alternative names

        • Villin 2 ezrin antibody
        • CVIL antibody
        • CVL antibody
        • Cytovillin 2 antibody
        • Cytovillin antibody
        • DKFZp762H157 antibody
        • Epididymis secretory protein Li 105 antibody
        • EZR antibody
        • EZRI_HUMAN antibody
        • Ezrin antibody
        • FLJ26216 antibody
        • HEL S 105 antibody
        • MGC1584 antibody
        • p81 antibody
        • VIL 2 antibody
        • VIL2 antibody
        • Villin 2 (ezrin) antibody
        • Villin 2 antibody
        • Villin-2 antibody
        • Villin2 antibody
        see all

      Images

      • Western blot - Anti-Ezrin antibody [EPR803] (ab133297)
        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.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)
        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.

      • Western blot - Anti-Ezrin antibody [EPR803] (ab133297)
        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 kDa



        Lanes 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.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ezrin antibody [EPR803] (ab133297)
        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.

      • Western blot - Anti-Ezrin antibody [EPR803] (ab133297)
        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)
        Anti-Ezrin antibody [EPR803] (ab133297)

      Protocols

      • Western blot protocols
      • Immunohistochemistry protocols

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
    • References (0)

      Publishing research using ab133297? Please let us know so that we can cite the reference in this datasheet.

      ab133297 has not yet been referenced specifically in any publications.

      Customer reviews and Q&As

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      There are currently no Customer reviews or Questions for ab133297.
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