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Alexa Fluor® 647 Mouse anti-p120 Catenin (pS268)
Alexa Fluor® 647 Mouse anti-p120 Catenin (pS268)

Analysis of p120 Catenin (pS268) in human epithelioid carcinoma.  After serum starvation overnight, Hela S3 cells (ATCC CCL 2.2) were either stimulated with 50 nM calyculin A (shaded histogram) for 30 minutes at 37°C or unstimulated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-p120 Catenin (pS268, Cat. No. 558559).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Analysis of p120 Catenin (pS268) in human epithelioid carcinoma.  After serum starvation overnight, Hela S3 cells (ATCC CCL 2.2) were either stimulated with 50 nM calyculin A (shaded histogram) for 30 minutes at 37°C or unstimulated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-p120 Catenin (pS268, Cat. No. 558559).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Phosflow™
Human (QC Testing), Mouse, Monkey, Dog (Reported)
Mouse IgG2b, κ
Phosphorylated Human p120 Catenin Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647149
Aqueous buffered solution containing BSA, protein stabilizer, 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558559 Rev. 2
Antibody Details
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9a.390

The membrane associated protein pp120 Src substrate (p120 catenin, p120cas) was identified as a tyrosine kinase substrate that is phosphorylated in Src-transformed cells.  It shares structural similarity with the Drosophila Armadillo protein and the vertebrate β-catenin and γ-catenin proteins in its 42-amino acid Arm domain.  p120 catenin is localized to the E-Cadherin/catenin cell adhesion complex.  Like β- and γ-catenin, p120 catenin directly associates with the cytoplasmic C-terminus of E-Cadherin via its Arm domain.  It exists as four isoforms that range in size from 90 to 115 kDa.  Expression of these isoforms is heterogeneous in human carcinomas, suggesting that altered expression contributes to malignancy.  Phosphorylation of multiple serine (S252, S268, S288, and S873), and threonine (T310 and T910) residues in p120 catenin may regulate its activity.  The S873 residue is phosphorylated after PKC activation, while the S268 site is dephosphorylated after PKC activation.  The latter residue is phosphorylated in vitro by p160 Rock.  S252 and T310 residues are phosphorylated in vitro by GSK3b.  Thus, p120 catenin function may be regulated in a complex manner through both serine and threonine phosphorylation.

The 9a.390 monoclonal antibody recognizes the phosphorylated S268 in the regulatory domain of p120 catenin.

558559 Rev. 2
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
558559 Rev.2
Citations & References
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Development References (2)

  1. Reynolds AB, Roczniak-Ferguson A. Emerging roles for p120-catenin in cell adhesion and cancer. Oncogene. 2004; 23:7947-7956. (Biology).
  2. Xia X, Brooks J, Campos-Gonzalez R, Reynolds AB. Serine and threonine phospho-specific antibodies to p120-catenin. Hybrid Hybridomics. 2004; 23:343-351. (Immunogen).
558559 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.