Analysis of Sox1 in H9 Embryonic Stem (ES) cells (left panel), H9-derived Neural Stem Cells (NSC) (middle panel), and H9-derived Neurons (right panel). H9 human ES cells (WiCell, Madison, WI), H9-derived NSC, and H9-derived neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), and permeabilized with BD™ Phosflow Perm buffer III (Cat. No. 558050). The ES and NSC were then blocked with 10% mouse serum (Sigma M9505). All cells were stained with matching concentrations of either PerCP-Cy5.5 Mouse IgG1, κ isotype control (dashed histograms, Cat. No. 550795) or PerCP-Cy5.5 Mouse Anti-Human Sox1 antibody (solid lines). Histograms were derived from gated events based on light scattering characteristics for the ES, NSC, and neurons, respectively. Flow cytometry was performed on a BD LSR™ II flow cytometry system. In the right panel, the brighter peak consists of undifferentiated NSC and glial cells that are staining positive for Sox1 and the dimmer peak consists of the neuronal population.
Analysis of Sox1 in H9 Embryonic Stem (ES) cells (left panel), H9-derived Neural Stem Cells (NSC) (middle panel), and H9-derived Neurons (right panel). H9 human ES cells (WiCell, Madison, WI), H9-derived NSC, and H9-derived neurons were harvested, fixed in BD Cytofix™ buffer (Cat. No. 554655), and permeabilized with BD™ Phosflow Perm buffer III (Cat. No. 558050). The ES and NSC were then blocked with 10% mouse serum (Sigma M9505). All cells were stained with matching concentrations of either PerCP-Cy5.5 Mouse IgG1, κ isotype control (dashed histograms, Cat. No. 550795) or PerCP-Cy5.5 Mouse Anti-Human Sox1 antibody (solid lines). Histograms were derived from gated events based on light scattering characteristics for the ES, NSC, and neurons, respectively. Flow cytometry was performed on a BD LSR™ II flow cytometry system. In the right panel, the brighter peak consists of undifferentiated NSC and glial cells that are staining positive for Sox1 and the dimmer peak consists of the neuronal population.
Product Details
BD Pharmingen™
Human (QC Testing)
Mouse IgG1, κ
Human Sox1 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_10694879
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
PerCP-Cy™5.5 Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
550795
561549 Rev. 1
Antibody Details
N23-844
The N23-844 monoclonal antibody reacts with human Sox1, a member of the SOX [SRY (sex determining region Y)-HMG-box] family of transcription factors. The encoded protein may act as a transcriptional activator after forming a protein complex with other proteins. It is one of the earliest transcription factors to be expressed in ectodermal cells committed to the neural fate. Sox1 is expressed in both embryonic and somatic neural stem and progenitor cells, and it is down regulated during neuronal differentiation in many neuronal subtypes.
561549 Rev. 1
Format Details
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.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
561549 Rev.1
Citations & References
Development References (4)
-
Kan L, Israsena N, Zhang Z, et al. Sox1 acts through multiple independent pathways to promote neurogenesis. Dev Biol. 2004; 15;26(2):580-594. (Biology). View Reference
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Malas S, Duthie SM, Mohri F, Lovell-Badge R, Episkopou V. Cloning and mapping of the human SOX1: a highly conserved gene expressed in the developing brain. Mamm Genome. 1997; 8(11):866-868. (Biology). View Reference
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Pevny LH, Sockanathan S, Placzek M, Lovell-Badge R. A role for SOX1 in neural determination. Development. 1998; 125(10):1967-1978. (Biology). View Reference
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Wilson M, Koopman P. Matching SOX: partner proteins and co-factors of the SOX family of transcriptional regulators. Curr Opin Genet Dev. 2002; 12(4):441-446. (Biology). View Reference
561549 Rev. 1
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.
