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Alexa Fluor® 647 Mouse anti-PTEN
Alexa Fluor® 647 Mouse anti-PTEN

Analysis of PTEN in human epithelioid carcinoma.  HeLa S3 cells (ATCC CCL 2.2) were either transfected with PTEN RNAi (open histogram) or untreated (shaded histogram).  The cells were fixed using BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C, then permeabilized usingBD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-PTEN.  Down-regulation of PTEN expression is evident in the RNAi-transfected cells.  Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.

Analysis of PTEN in human epithelioid carcinoma.  HeLa S3 cells (ATCC CCL 2.2) were either transfected with PTEN RNAi (open histogram) or untreated (shaded histogram).  The cells were fixed using BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C, then permeabilized usingBD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-PTEN.  Down-regulation of PTEN expression is evident in the RNAi-transfected cells.  Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.

Product Details
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BD Phosflow™
MMAC1, TEP1
Human (QC Testing), Mouse (Tested in Development), Rat, Dog (Reported)
Mouse BALB/c IgG1, κ
Human C-terminal PTEN Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645437
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. An isotype control should be used at the same concentration as the antibody of interest.
  2. 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.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. Alexa Fluor® is a registered 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. 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).
  9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560003 Rev. 4
Antibody Details
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A2B1

Cancer can develop when cells escape normal growth control mechanisms through mutations in proto-oncogenes or tumor suppressor genes.  A characteristic of most oncogene and tumor suppressor gene products is that they are components of signal transduction pathways that are essential for maintaining cellular homeostasis.  PTEN (phosphatase and tensin homolog), also known as MMAC1 (mutated in multiple advanced cancers 1), is a tumor suppressor gene that is mutated at high frequency in multiple tumor types.  The protein encoded by PTEN is a phosphatase that preferentially dephosphorylates phosphoinositide substrates.  It is believed that a mechanism by which PTEN mutations cause tumors is the loss of its negative control on the phosphoinositide 3-kinase signaling pathway that regulates cell growth and survival.  PTEN also plays a role in the maintenance of hematopoietic stem cells.

The A2B1 monoclonal antibody recognizes PTEN, regardless of phosphorylation status.

The specificity of this antibody conjugate for flow cytometric analysis was validated by confirming that RNA-mediated interference (RNAi) of the specific protein reduced the staining of the cells (see figure).  Furthermore, the capacity of the RNAi to down-regulate the expression of the relevant protein was confirmed by western blot analysis (data not shown).

560003 Rev. 4
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
560003 Rev.4
Citations & References
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Development References (8)

  1. Cantley LC, Neel BG. New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway. Proc Natl Acad Sci U S A. 1999; 96(8):4240-4245. (Biology). View Reference
  2. Goberdhan DCI, Wilson C. PTEN: tumour suppressor, multifunctional growth regulator and more. Hum Mol Genet. 2003; 12(2):R238-R248. (Biology).
  3. Levine RA, Forest T, Smith C. Tumor suppressor PTEN is mutated in canine osteosarcoma cell lines and tumors. Vet Pathol. 2002; 39:372-378. (Clone-specific: Immunohistochemistry).
  4. Raftopoulou M, Etienne-Manneville S, Self A, Nicholls S, Hall A. Regulation of cell migration by the C2 domain of the tumor suppressor PTEN. Science. 2004; 303:1179-1181. (Biology).
  5. Steck PA, Pershouse MA, Jasser SA, et al. Identification of a candidate tumour suppressor gene, MMAC1 at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet. 1997; 15(4):356-362. (Biology). View Reference
  6. Wu R-C, Li X, Schönthal AH. Transcriptional activation of p21WAF1 by PTEN/MMAC1 tumor suppressor. Mol Cell Biochem. 2000; 203:59-71. (Immunogen: Western blot).
  7. Yilmaz OH, Valez R, Theisen BK, et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature. 2006; 441:475-482. (Biology).
  8. Zhang J, Grindley JC, Yin T, et al. PTEN maintains haematopoietic stem cells and acts in lineage choice and leukaemia prevention. Nature. 2006; 441:518-522. (Biology).
View All (8) View Less
560003 Rev. 4

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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.