Flow cytometric analysis of SOX9 expression in human hepatocellular carcinoma and embryonic stem cell lines. Hep G2 cells (ATCC HB-8065, Left Panel) and H9 cells (WiCell, Madison, WI; Right Panel) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No.557732; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Sox9 (Cat. No. 565493; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact Hep G2 or H9 cells, respectively. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Flow cytometric analysis of SOX9 expression in human hepatocellular carcinoma and embryonic stem cell lines. Hep G2 cells (ATCC HB-8065, Left Panel) and H9 cells (WiCell, Madison, WI; Right Panel) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No.557732; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Sox9 (Cat. No. 565493; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact Hep G2 or H9 cells, respectively. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
BD Pharmingen™
SOX-9, SOX9
Human (QC Testing), Rat (Tested in Development)
Mouse IgG1, κ
Human Sox9 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Not Recommended)
0.2 mg/ml
AB_2739262
Aqueous buffered solution containing 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.
Recommended Assay Procedures
For Bioimaging and Immunofluorescence applications we recommend that researchers consider Purified Mouse Anti-Sox 9 (Clone T32-668; Cat. No. 565492). For additional information please reference http://www.bdbiosciences.com/resources/index.jsp
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557732
565493 Rev. 1
Antibody Details
T32-668
The T32-668 monoclonal antibody specifically binds to Sox9, a member of the SRY-related HMG-box (SOX) family of transcription factors. Sox9 plays a key role in skeletal development and sex determination. Mutation of Sox9 has been associated with the human skeletal dysmorphology syndrome campomelic dysplasia, which is often characterized by both skeletal abnormalities and sex reversal. Expression of Sox9 has been observed in a variety of tissues and organs, including cartilage, testis, central neural system, lung, pancreas, intestinal stem cells, and heart.
565493 Rev. 1
Format Details
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
565493 Rev.1
Citations & References
Development References (5)
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Furuyama K, Kawaguchi Y, Akiyama H, et al. Continuous cell supply from a Sox9-expressing progenitor zone in adult liver, exocrine pancreas and intestine. Nat Genet. 2011; 43(1):34-41. (Biology). View Reference
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Guo W, Keckesova Z, Donaher JL, et al. Slug and Sox9 cooperatively determine the mammary stem cell state. Cell. 2012; 148(5):1015-1028. (Biology). View Reference
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Kent J, Wheatley SC, Andrews JE, Sinclair AH, Koopman P. A male-specific role for SOX9 in vertebrate sex determination. Development. 1996; 122(9):2813-2822. (Biology). View Reference
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Stolt CC, Lommes P, Sock E, Chaboissier MC, Schedl A, Wegner M. The Sox9 transcription factor determines glial fate choice in the developing spinal cord. Genes Dev. 2003; 17(13):1677-1689. (Biology). View Reference
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Wright E, Hargrave MR, Christiansen J, et al. The Sry-related gene Sox9 is expressed during chondrogenesis in mouse embryos. Nat Genet. 1995; 9(1):15-20. (Biology). View Reference
565493 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.
