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Alexa Fluor® 647 Mouse anti-Oct3/4
Alexa Fluor® 647 Mouse anti-Oct3/4

Flow cytometric analysis of Alexa Fluor® 647 anti-Oct3/4 in H9 cells.  H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were grown on BD Matrigel™ hESC-qualified matrix (Cat. No. 354277).  Cells were dissociated into a single cell suspension and fixed (BD Cytofix™ buffer, Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885), and then stained with either Alexa Fluor® 647 Mouse anti-Oct3/4 (solid line) or Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557783, dashed line).  Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. This antibody also works in BD™ Phosflow Perm Buffer II and III.

Flow cytometric analysis of Alexa Fluor® 647 anti-Oct3/4 in H9 cells.  H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were grown on BD Matrigel™ hESC-qualified matrix (Cat. No. 354277).  Cells were dissociated into a single cell suspension and fixed (BD Cytofix™ buffer, Cat. No. 554655), permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885), and then stained with either Alexa Fluor® 647 Mouse anti-Oct3/4 (solid line) or Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557783, dashed line).  Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. This antibody also works in BD™ Phosflow Perm Buffer II and III.

Product Details
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BD Pharmingen™
Oct3, OTF3, Oct4, OTF4, POU5F1
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Mouse Oct3 aa. 252-372 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645318
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  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® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. 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.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  9. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
560329 Rev. 4
Antibody Details
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40/Oct-3

Development of a multicellular organism from a single fertilized cell is regulated by the coordinated activity of DNA transcription factors.  Oct3/4, a member of the POU family of transcription factors, functions in pluripotent cells of early embryonic stem cell (ES) lines and embryonal carcinomas (EC).  Other members of the POU family include Oct1, Oct2, Pit-1, and unc-86.  The POU domain, a 150-amino acid region that determines binding specificity, is conserved among these proteins and consists of 3 subdomains: POU-specific A and B subdomains and a homeobox-like subdomain.  Oct3/4 is expressed in undifferentiated cells, but is lost as cells are induced to differentiate.  Oct3/4 is not expressed in adult tissues.  The interaction of Oct3/4 with SOX2, another embryonic transcription factor, produces an active complex that regulates expression of genes such as Nanog, UTF1, and FGF4.  Although Oct3/4 is specifically phosphorylated on serine residues, this modification is not required for DNA binding, but may affect its transactivation potential.  Thus, Oct3/4 is a transcription factor that plays an important role in determining early steps of embryogenesis and differentiation.

560329 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
560329 Rev.4
Citations & References
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Development References (6)

  1. Nishimoto M, Fukushima A, Okuda A, Muramatsu M. The gene for the embryonic stem cell coactivator UTF1 carries a regulatory element which selectively interacts with a complex composed of Oct-3/4 and Sox-2. Mol Cell Biol. 1999; 19(8):5453-5465. (Biology). View Reference
  2. Okamoto K, Okazawa H, Okuda A, Sakai M, Muramatsu M, Hamada H. A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells. Cell. 1990; 60(3):461-472. (Biology). View Reference
  3. Pan G, Thomson JA. Nanog and transcriptional networks in embryonic stem cell pluripotency. Cell Res. 2007; 17:42-49. (Biology). View Reference
  4. Rosfjord E, Scholtz B, Lewis R, Rizzino A. Phosphorylation and DNA binding of the octamer binding transcription factor Oct-3. Biochem Biophys Res Commun. 1995; 212(3):847-853. (Biology). View Reference
  5. Vigano MA, Staudt LM. Transcriptional activation by Oct-3: evidence for a specific role of the POU-specific domain in mediating functional interaction with Oct-1. Nucleic Acids Res. 1996; 24(11):2112-2118. (Biology). View Reference
  6. Yuan H, Corbi N, Basilico C, Dailey L. Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3. Genes Dev. 1995; 9(21):2635-2645. (Biology). View Reference
View All (6) View Less
560329 Rev. 4

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.