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PerCP-Cy™5.5 Mouse anti-Oct3/4
PerCP-Cy™5.5 Mouse anti-Oct3/4
Flow cytometric analysis of PerCP-Cy5.5 Mouse anti-Oct3/4 in human embryonic stem cells.  H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were fixed with BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD™ Phosflow Perm/Wash buffer I (Cat. No. 557885), and then stained with either PerCP-Cy5.5 Mouse anti-Oct3/4 (Cat. No. 560794, solid line) or PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (Cat. No. 552834, dashed line).  Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. This antibody will also work in BD™ Phosflow Perm II and III buffers.
Flow cytometric analysis of PerCP-Cy5.5 Mouse anti-Oct3/4 in human embryonic stem cells.  H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were fixed with BD Cytofix™ buffer (Cat. No. 554655), permeabilized with BD™ Phosflow Perm/Wash buffer I (Cat. No. 557885), and then stained with either PerCP-Cy5.5 Mouse anti-Oct3/4 (Cat. No. 560794, solid line) or PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (Cat. No. 552834, dashed line).  Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system. This antibody will also work in BD™ Phosflow Perm II and III buffers.
Product Details
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BD Pharmingen™
Oct3, OTF3, Oct4, OTF4, POU5F1
Human (QC Testing), Mouse (Reported)
Mouse IgG1, κ
Mouse Oct3 aa. 252-372 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1937313
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
  5. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  6. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  8. 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.
  9. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  10. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560794 Rev. 2
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.

560794 Rev. 2
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
560794 Rev.2
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
560794 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.