Skip to main content Skip to navigation
BV421 Mouse Anti-E-Cadherin
BV421 Mouse Anti-E-Cadherin
Flow cytometric analysis of E-cadherin expression in a human breast carcinoma cell line (left panel). Cells from the human MCF-7 (Breast adenocarcinoma, ATCC HTB-22) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 563464, dashed line histogram) or BD Horizon BV421 Mouse Anti-E-Cadherin antibody (Cat. No. 564186). Histograms were derived from gated events with the forward and side light-scattering characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometry System. Western blot analysis of E-Cadherin (center panel) using Purified Mouse Anti-E-Cadherin. E-Cadherin is observable at 120kDa. A431 lysate (ATCC CRL-1555; human epithelial carcinoma, left blot) was blotted at 1:10000 & 1:20000 (Lanes 1 & 2 respectively; 30 second exposure). 293F control lysate (center left blot) was blotted at 1:250 & 1:500 (Lanes 1 & 2 respectively; 30 second exposure).  293F cells transfected with human E-Cadherin (CDH1, center right blot) was blotted at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure). 293 cells transfected with human P-Cadherin (CDH3, right blot) was blotted using Purified Mouse Anti-E-Cadherin (Cat. No. 610181/610182) at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure).
BV421 Mouse Anti-E-Cadherin

Immunofluorescent analysis of E-Cadherin expression. MCF7 (ATCC HTB-22) cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), and blocked with 5% goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in PBS. Cells were stained with BD Horizon™ BV421 Mouse Anti-E-Cadherin antibody (Cat. No. 564186, pseudo-colored green) and Alexa Fluor® 555 Mouse Anti-Human Ki-67 antibody (Cat. No. 558617, pseudo-colored red). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored blue). Images were captured on a standard four laser confocal microscope. Original magnification, 40x.

Flow cytometric analysis of E-cadherin expression in a human breast carcinoma cell line (left panel). Cells from the human MCF-7 (Breast adenocarcinoma, ATCC HTB-22) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) and stained with either BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 563464, dashed line histogram) or BD Horizon BV421 Mouse Anti-E-Cadherin antibody (Cat. No. 564186). Histograms were derived from gated events with the forward and side light-scattering characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometry System. Western blot analysis of E-Cadherin (center panel) using Purified Mouse Anti-E-Cadherin. E-Cadherin is observable at 120kDa. A431 lysate (ATCC CRL-1555; human epithelial carcinoma, left blot) was blotted at 1:10000 & 1:20000 (Lanes 1 & 2 respectively; 30 second exposure). 293F control lysate (center left blot) was blotted at 1:250 & 1:500 (Lanes 1 & 2 respectively; 30 second exposure).  293F cells transfected with human E-Cadherin (CDH1, center right blot) was blotted at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure). 293 cells transfected with human P-Cadherin (CDH3, right blot) was blotted using Purified Mouse Anti-E-Cadherin (Cat. No. 610181/610182) at 1:2500 & 1: 5000 (Lanes 1 & 2 respectively; 5 second exposure).

Immunofluorescent analysis of E-Cadherin expression. MCF7 (ATCC HTB-22) cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), and blocked with 5% goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in PBS. Cells were stained with BD Horizon™ BV421 Mouse Anti-E-Cadherin antibody (Cat. No. 564186, pseudo-colored green) and Alexa Fluor® 555 Mouse Anti-Human Ki-67 antibody (Cat. No. 558617, pseudo-colored red). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored blue). Images were captured on a standard four laser confocal microscope. Original magnification, 40x.

Product Details
Down Arrow Up Arrow


BD Horizon™
CD324; CDH1; CADH1; Cadherin-1; ECAD; CDHE; Arc-1; LCAM; UVO; Uvomorulin
Human (QC Testing), Mouse, Rat, Dog (Tested in Development)
Mouse IgG2a, κ
Human E-Cadherin C-terminal Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Immunocytochemistry, Immunofluorescence (Tested During Development)
5 µl
AB_2738654
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  10. Triton is a trademark of the Dow Chemical Company.
  11. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564186 Rev. 3
Antibody Details
Down Arrow Up Arrow
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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564186 Rev. 3
Format Details
Down Arrow Up Arrow
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV421
Violet 405 nm
407 nm
423 nm
564186 Rev.3
Citations & References
Down Arrow Up Arrow

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
View All (5) View Less
564186 Rev. 3

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.