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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.
BD Pharmingen™ Alexa Fluor® 647 Mouse anti-Oct3/4
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- 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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
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.
Development References (6)
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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
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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
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Pan G, Thomson JA. Nanog and transcriptional networks in embryonic stem cell pluripotency. Cell Res. 2007; 17:42-49. (Biology). View Reference
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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
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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
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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
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.