Anti-Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) antibody [EP2122Y] (ab76247)

Overview

  • Product name
    Anti-Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) antibody [EP2122Y]
  • Description
    Rabbit monoclonal [EP2122Y] to Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558)
  • Specificity
    ab76247 will detect Ezrin, Radixin and Moesin. This antibody detects the phosphorylated target and not the non-phosphorylated epitope.
  • Tested applications
    Suitable for: WB, IP, IHC-P, Dot blotmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Ezrin (pThr567)/ Radixin (pThr564)/ Moesin (pThr558). A synthetic peptide corresponding to residues surrounding Thr567 (Ezrin), Thr564 (Radixin) and Thr558 (Moesin) of human Ezrin.

  • Positive control
    • IHC: human breast adenocarcinoma IF: HeLa cells Dot Blot: Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) phospho peptide and Non-phospho peptide.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76247 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 1/1000. Detects a band of approximately 75-81 kDa (predicted molecular weight: 69 kDa).
IP 1/40.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Dot blot 1/1000.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Images

    • All lanes : Anti-Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) antibody [EP2122Y] (ab76247) at 1/1000 dilution

      Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with 1 µM Staurosporine for 5 hours whole cell lysates.

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 69 kDa


      Exposure time : 30 seconds

      Blocking and diluting buffer: 2% BSA/TBST.

      Staurosporine reduces ERM phosphorylation (PMID:26555866 and 23209304)

    • Immunohistochemical analysis of paraffin-embedded human breast adenocarcinoma with ab76247 at 1/100 dilution.

    • All lanes : Anti-Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) antibody [EP2122Y] (ab76247) at 1/1000 dilution

      Lane 1 : 293 (Human embryonic kidney epithelial cell) treated with Calyculin A whole cell lysates)
      Lane 2 : 293 (Human embryonic kidney epithelial cell) treated with Calyculin A whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.
      Lane 3 : 293 (Human embryonic kidney epithelial cell) treated with Calyculin A whole cell lysates. Then the membrane was incubated with Lambda phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 69 kDa


      Exposure time : 1 second

      Blocking and diluting buffer and concentration: 5% NFDM/TBST
      This antibody detects two bands at 75-81KDa.

    • All lanes : Anti-Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) antibody [EP2122Y] (ab76247) at 1/5000 dilution

      Lane 1 : 293 (Human embryonic kidney epithelial cell) treated with Calyculin A whole cell lysates
      Lane 2 : 293 (Human embryonic kidney epithelial cell) treated with Calyculin A whole cell lysates. Then the membrane was incubated with Lambda phosphatase.
      Lane 3 : 293 (Human embryonic kidney epithelial cell) treated with Calyculin A whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 69 kDa


      Exposure time : 3 minutes

      Blocking and diluting buffer: 5% NFDM/TBST

    • Dot blot analysis of Ezrin (pThr567)/ Radixin (pThr564)/ Moesin (pThr558) phospho peptide (Lane 1), Non-phospho peptide (Lane 2), labelling Ezrin (phospho Thr567)/ Radixin (phospho Thr564)/ Moesin (phospho Thr558) with ab76247 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

      Blocking and diluting buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

      The sequence of non-phospho peptide: CGRDKYKTLRQIR.
      The sequence of phospho peptide: CGRDKYK-pT-LRQIR.

    References

    This product has been referenced in:
    • Lee A  et al. Vitamin D Proliferates Vaginal Epithelium through RhoA Expression in Postmenopausal Atrophic Vagina tissue. Mol Cells 40:677-684 (2017). WB ; Human . Read more (PubMed: 28843271) »
    • Luján P  et al. PRL-3 disrupts epithelial architecture by altering the post-mitotic midbody position. J Cell Sci 129:4130-4142 (2016). Read more (PubMed: 27656108) »

    See all 4 Publications for this product

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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