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PE Mouse anti-Ezrin (pY353)
PE Mouse anti-Ezrin (pY353)

Analysis of Ezrin (pY353) in human chronic myelogenous leukemia.  Serum-starved K-562 cells (ATCC CCL-243) were either treated with 1 mM Pervanadate (Sigma Cat. No. S6508) at 37°C for 20 minutes (shaded histogram) or untreated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-Ezrin (pY353, Cat. No. 558601).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Analysis of Ezrin (pY353) in human chronic myelogenous leukemia.  Serum-starved K-562 cells (ATCC CCL-243) were either treated with 1 mM Pervanadate (Sigma Cat. No. S6508) at 37°C for 20 minutes (shaded histogram) or untreated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-Ezrin (pY353, Cat. No. 558601).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Phosflow™
Human (QC Testing)
Mouse IgG1, κ
Phosphorylated Human Ezrin Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647206
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558601 Rev. 2
Antibody Details
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I66-386

Ezrin is a member of the ERM (ezrin-radixin-moesin) family of proteins that function as crosslinkers between the actin cytoskeleton and the plasma membrane.  Ezrin is a substrate for the tyrosine kinase epidermal (EGF) and hepatocyte growth factor (HGF) receptors.  Upon EGF or HGF stimulation, ezrin is phosphorylated on tyrosine 353 (Y353), which is required for its interaction with phosphatidylinositol 3 kinase p85 subunit and activation of Akt.

The I66-386 monoclonal antibody recognizes the phosphorylated Y353 of human ezrin.

558601 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
558601 Rev.2
Citations & References
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Development References (2)

  1. Gautreau A, Poullet P, Louvard D, Arpin M. Ezrin, a plasma membrane–microfilament linker, signals cell survival through the phosphatidylinositol 3-kinase/Akt pathway. Proc Natl Acad Sci U S A. 1999; 96:7300-7305. (Biology). View Reference
  2. Krieg J, Hunter T. Identification of the two major epidermal growth factor-induced tyrosine phosphorylation sites in the microvillar core protein ezrin. J Biol Chem. 1992; 267(27):19258-19265. (Biology).
558601 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.