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PE Mouse Anti-Human CD324 (E-Cadherin)
PE Mouse Anti-Human CD324 (E-Cadherin)

Flow Cytometric Analysis of CD324 (E-cadherin) on human breast adenocarcinoma (left) and human embryonic stem (ES) cells (right). Human breast adenocarcinoma cell line MCF-7 (ATCC, HTB-22) and H9 human ES Cells (WiCell, Madison, WI) were harvested with Cell Dissociation Buffer (Life Technologies). Please note that the epitope is sensitive to Accutase™ detachment. Cells were stained with either PE Mouse IgG1, k isotype control (Cat. No. 554680; dashed line) or PE Mouse anti-human CD324 antibody (Cat. No. 562870; solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of MCF-7 or human ES cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. Staining with this antibody can also performed on cells that have been fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885). However, the brightness of the staining decreases after cell fixation and permeabilization.

Flow Cytometric Analysis of CD324 (E-cadherin) on human breast adenocarcinoma (left) and human embryonic stem (ES) cells (right). Human breast adenocarcinoma cell line MCF-7 (ATCC, HTB-22) and H9 human ES Cells (WiCell, Madison, WI) were harvested with Cell Dissociation Buffer (Life Technologies). Please note that the epitope is sensitive to Accutase™ detachment. Cells were stained with either PE Mouse IgG1, k isotype control (Cat. No. 554680; dashed line) or PE Mouse anti-human CD324 antibody (Cat. No. 562870; solid line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of MCF-7 or human ES cells. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. Staining with this antibody can also performed on cells that have been fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885). However, the brightness of the staining decreases after cell fixation and permeabilization.

Product Details
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BD Pharmingen™
E-cadherin; CD324; CDH1; CADH1; Cadherin-1; ECAD; CDHE; Arc-1; LCAM; UVO
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Breast Tumor Cell Line
Flow cytometry (Routinely Tested)
5 µl
VIII 80167
999
AB_2737854
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

In addition to standard cell surface staining of live cells, this antibody conjugate may also be used to stain cells that have been fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885). Because the brightness of the staining decreases after cell fixation and permeabilization, the reagent titer may need to be adjusted.

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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. All other brands are trademarks of their respective owners.
562870 Rev. 1
Antibody Details
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67A4

The 67A4 monoclonal antibody specifically recognizes the extracellular domain of human E-Cadherin (CD324). 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-to-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 their invasiveness. Thus, loss of expression or function of E-Cadherin appears to be an important step in tumorigenic progression. Pluripotent stem cells express E-Cadherin. Upon differentiation, an epithelial to mesenchymal transition results in the loss of E-cadherin expression and a gain in the expression of N-cadherin.

562870 Rev. 1
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.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
562870 Rev.1
Citations & References
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Development References (5)

  1. Behrens J, Vakaet L, Friis R, et al. Loss of epithelial differentiation and gain of invasiveness correlates with tyrosine phosphorylation of the E-cadherin/beta-catenin complex in cells transformed with a temperature-sensitive v-SRC gene.. J Cell Biol. 1993; 120(3):757-66. (Biology). View Reference
  2. Bühring HJ, Müller T, Herbst R, et al. The adhesion molecule E-cadherin and a surface antigen recognized by the antibody 9C4 are selectively expressed on erythroid cells of defined maturational stages.. Leukemia. 1996; 10(1):106-16. (Clone-specific: Flow cytometry). View Reference
  3. Cepek KL, Shaw SK, Parker CM, et al. Adhesion between epithelial cells and T lymphocytes mediated by E-cadherin and the alpha E beta 7 integrin.. Nature. 1994; 372(6502):190-3. (Biology). View Reference
  4. D'Amour KA, Agulnick AD, Eliazer S, Kelly OG, Kroon E, Baetge EE. Efficient differentiation of human embryonic stem cells to definitive endoderm.. Nat Biotechnol. 2005; 23(12):1534-41. (Biology). View Reference
  5. Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis.. Development. 1988; 102(4):639-55. (Biology). View Reference
View All (5) View Less
562870 Rev. 1

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.