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Alexa Fluor® 647 Mouse Anti-Human CD325
Alexa Fluor® 647 Mouse Anti-Human CD325

Flow cytometric analysis of CD325 (N-Cadherin) on transformed human HeLa cells. Cells from the HeLa (cervical adenocarcinoma, ATCC CCL-2.2) cell line were harvested with a cell dissociation buffer without trypsin (please note, the CD325 epitope is sensitive to trypsin). The cells were then washed and stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714, dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human CD325 antibody (Cat. No. 563434; solid line histogram). The histograms were derived from gated events based on the forward and side light-scatter characteristics of viable HeLa cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD325 (N-Cadherin) on transformed human HeLa cells. Cells from the HeLa (cervical adenocarcinoma, ATCC CCL-2.2) cell line were harvested with a cell dissociation buffer without trypsin (please note, the CD325 epitope is sensitive to trypsin). The cells were then washed and stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714, dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human CD325 antibody (Cat. No. 563434; solid line histogram). The histograms were derived from gated events based on the forward and side light-scatter characteristics of viable HeLa cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
Cadherin-2, N-Cadherin; NCAD; CDHM; CDH2
Human (QC Testing)
Mouse IgG1, κ
Human extracellular N-Cadherin domain Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
1000
AB_2738202
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was 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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563434 Rev. 1
Antibody Details
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8C11

The 8C11 monoclonal antibody recognizes the extracellular domain of human N-Cadherin (CD325). Cadherins are a family of Ca2+ -dependent intercellular adhesion molecules that play a central role in controlling morphogenetic movements during development. Their function is regulated by association with the actin cytoskeleton by a complex of cytoplasmic proteins called the catenins (α, β, γ). Members of the cadherin family include P-cadherin , E-cadherin (uvomorulin), N-cadherin (neural cadherin), R-cadherin, cadherin 5, L-CAM, and EP-cadherin. N-cadherin mRNA is found at elevated levels in brain and heart and at a much lower level in liver. Mechanisms such as mRNA expression, cytokine modulation, and protease-mediated turnover modulate N-cadherin protein levels during development. In addition, N-cadherin function is indirectly regulated by endogenous kinases and phosphatases. Tyrosine phosphorylation of β-catenin complexed with N-cadherin results in dissociation of N-cadherin from actin. However, N-cadherin also interacts with a PTP1B-like phosphatase that dephosphorylates β-catenin and promotes N-cadherin/actin association. Thus, N-cadherin is an integral adhesion molecule whose function is regulated by protein-protein interactions and phosphorylation/dephosphorylation events.

563434 Rev. 1
Format Details
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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.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563434 Rev.1
Citations & References
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Development References (3)

  1. Knudsen KA, Soler AP, Johnson KR, Wheelock MJ. Interaction of alpha-actinin with the cadherin/catenin cell-cell adhesion complex via alpha-catenin. J Cell Biol. 1995; 130:66-77. (Biology). View Reference
  2. Puch S, Armeanu S, Kibler C, et al. N-cadherin is developmentally regulated and functionally involved in early hematopoietic cell differentiation. J Cell Sci. 2001; 114(8):1567-1577. (Clone-specific: Flow cytometry). View Reference
  3. Wein F, Pietsch L, Saffrich R, et al. N-Cadherin is expressed on human hematopoietic progenitor cells and mediates interaction with human mesenchymal stromal cells. Stem Cell Res. 2010; 4(2):129-139. (Clone-specific: Flow cytometry). View Reference
563434 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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.