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PE Mouse anti-E-Cadherin
PE Mouse anti-E-Cadherin
Left Image: Flow cytometric analysis of E-cadherin expressed in a human breast carcinoma cell line. MCF-7 cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with matching concentrations of either PE Mouse IgG2a, κ Isotype Control (Cat. No. 558595, dashed line histogram) or PE Mouse Anti-E-Cadherin antibody (Cat. No. 562526). Histograms were derived from gated events with the forward and side light-scattering characteristics of intact cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System. Right Image: Western blot analysis of E-Cadherin using Purified Mouse Anti-E-Cadherin (Cat. No. 610181/610182). E-Cadherin is observable at 120kDa. Left Panel: A431 lysate (ATCC CRL-1555; human epithelial carcinoma) was blotted at 1:10000 & 1:20000 (Lanes 1 & 2 respectively; 30 second exposure). Middle Left Panel: 293F control lysate was blotted at 1:250 & 1:500 (Lanes 1 & 2 respectively; 30 second exposure). Middle Right Panel: 293F cells transfected with human E-Cadherin (CDH1) was blotted at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure). Right Panel: 293 cells transfected with human P-Cadherin (CDH3) was blotted using Purified Mouse Anti-E-Cadherin (Cat. No. 610181/610182) at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure).
Left Image: Flow cytometric analysis of E-cadherin expressed in a human breast carcinoma cell line. MCF-7 cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with matching concentrations of either PE Mouse IgG2a, κ Isotype Control (Cat. No. 558595, dashed line histogram) or PE Mouse Anti-E-Cadherin antibody (Cat. No. 562526). Histograms were derived from gated events with the forward and side light-scattering characteristics of intact cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System. Right Image: Western blot analysis of E-Cadherin using Purified Mouse Anti-E-Cadherin (Cat. No. 610181/610182). E-Cadherin is observable at 120kDa. Left Panel: A431 lysate (ATCC CRL-1555; human epithelial carcinoma) was blotted at 1:10000 & 1:20000 (Lanes 1 & 2 respectively; 30 second exposure). Middle Left Panel: 293F control lysate was blotted at 1:250 & 1:500 (Lanes 1 & 2 respectively; 30 second exposure). Middle Right Panel: 293F cells transfected with human E-Cadherin (CDH1) was blotted at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure). Right Panel: 293 cells transfected with human P-Cadherin (CDH3) was blotted using Purified Mouse Anti-E-Cadherin (Cat. No. 610181/610182) at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure).
Product Details
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BD Pharmingen™
CD324; CDH1; CADH1; Cadherin-1; ECAD; CDHE; Arc-1; LCAM; UVO; Uvomorulin
Human (QC Testing), Mouse, Rat, Dog (Reactivity Confirmed in Development)
Mouse IgG2a, κ
Human E-Cadherin C-terminal Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_11153868
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. 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.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
562526 Rev. 2
Antibody Details
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36/E-Cadherin

E-Cadherin is a 120-kDa transmembrane glycoprotein that is localized in the adherens junctions of epithelial cells.  There it interacts with the cytoskeleton through the associated cytoplasmic catenin proteins.  In addition to being a calcium-dependent adhesion molecule, E-Cadherin is also a critical regulator of epithelial junction formation.  Its association with catenins is necessary for cell-cell adhesion.  These E-cadherin/catenin complexes associate with cortical actin bundles at both the zonula adherens and the lateral adhesion plaques.  Tyrosine phosphorylation can disrupt these complexes, leading to changes in cell adhesion properties.  E-Cadherin expression is often down-regulated in highly invasive, poorly differentiated carcinomas.  Increased expression of E-Cadherin in these cells reduces invasiveness.  Thus, loss of expression or function of E-Cadherin appears to be an important step in tumorigenic progression. The 36/E-Cadherin monoclonal antibody recognizes the cytoplasmic domain of E-Cadherin, regardless of phosphorylation status.  The peptide immunogen was generated from human E-Cadherin aa. 735-883.

Note: Investigators are advised that this antibody has some degree of cross-reactivity to P-Cadherin.

562526 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
562526 Rev.2
Citations & References
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Development References (5)

  1. Jaksits S, Kriehuber E, Charbonnier AS, Rappersberger K, Stingl G, Maurer D. CD34+ cell-derived CD14+ precursor cells develop into Langerhans cells in a TGF-beta 1-dependent manner. J Immunol. 1999; 163(9):4869-4877. (Clone-specific: Immunofluorescence, Western blot). View Reference
  2. Miyoshi K, Shillingford JM, Smith GH, et al. Signal transducer and activator of transcription (Stat) 5 controls the proliferation and differentiation of mammary alveolar epithelium. J Cell Biol. 2001; 155(4):531-542. (Clone-specific: Immunohistochemistry). View Reference
  3. Sheibani N, Sorenson CM, Frazier WA. Differential modulation of cadherin-mediated cell-cell adhesion by platelet endothelial cell adhesion molecule-1 isoforms through activation of extracellular regulated kinases. Mol Biol Cell. 2000; 11(8):2793-2802. (Clone-specific: Immunofluorescence, Western blot). View Reference
  4. Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis.. Development. 1988; 102(4):639-55. (Biology). View Reference
  5. Weng Z, Xin M, Pablo L, et al. Protection against anoikis and down-regulation of cadherin expression by a regulatable beta-catenin protein. J Biol Chem. 2002; 277(21):18677-18686. (Clone-specific: Immunofluorescence, Immunoprecipitation). View Reference
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562526 Rev. 2

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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.