Alexa Fluor® 647 Mouse Anti-Human CD324 (E-Cadherin)

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BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human CD324 (E-Cadherin)

Name: Alexa Fluor® 647 Mouse Anti-Human CD324 (E-Cadherin)
Brand: BD Pharmingen™
SKU: 563571

Clone 67A4

(RUO)

Flow cytometric analysis of CD324 (E-cadherin) expression in human embryonic stem (ES) cells. 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. The cells were stained with either Alexa Fluor® 647 Mouse anti-Human CD324 antibody (Cat. No. 563571, solid line) or Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714, dashed line) at matched concentrations. Histograms were derived from gated events based on light scattering characteristics of H9 human ES cells. Flow cytometry was performed on a BD LSRFortessa™ Flow Cytometry System.

Immunofluorescent Staining of CD324 (E-Cadherin) on human embryonic stem (ES) cells. H9 human ES cells cells (WiCell, Madison, WI) grown in mTESR™1 media (Stem Cell Technologies) were fixed with BD Cytofix™ fixation buffer (Cat. No. 554655). Cells were stained with Alexa Fluor® 647 Mouse Anti-Human CD324 (E-Cadherin) monoclonal antibody (Cat. No. 563571, pseudo-colored red) at 5 µg/mL. Counter-staining was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD AttoVision™ Software.

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Product Details

Brand: BD Pharmingen™

Alternative Name: E-cadherin; CD324; CDH1; CADH1; Cadherin-1; ECAD; CDHE; Arc-1; LCAM; UVO

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human Breast Tumor Cell Line

Application: Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)

Vol. Per Test: 5 µl

Workshop Number: VIII 80167

Entrez Gene ID: 999

RRID: AB_2738283

Storage Buffer: Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.

Regulatory Status: 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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

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).
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
An isotype control should be used at the same concentration as the antibody of interest.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Accutase is a registered trademark of Innovative Cell Technologies, Inc.
mTESR™1 is a trademark of StemCell Technologies.

Companion Products

Fixation Buffer RUO

Size 100 mL Cat No. 554655

Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO

Size 100 Tests Cat No. 557714

563571 Rev. 1

Antibody Details

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.

563571 Rev. 1

Format Details

Alexa Fluor™ 647

Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: Alexa Fluor™ 647

Excitation Source: Red 627-640 nm

Excitation Max: 653 nm

Emission Max: 669 nm

563571 Rev.1

Citations & References

Development References (5)

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
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). View Reference
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
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
Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis.. Development. 1988; 102(4):639-55. (Biology). View Reference

View All (5)

563571 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.