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Flow cytometric analysis of Alexa Fluor® 647 Mouse Anti-SSEA-4 on human embryonic stem cells. H9 human embryonic stem (ES) cells (WiCell, Madison, WI) were stained with either Alexa Fluor® 647 Mouse Anti-SSEA-4 (solid line) or Alexa Fluor® 647 mouse IgG3, k Isotype Control (Cat. No. 560803, dashed line). Flow cytometry was performed on a BD™ LSR II flow cytometry system.
BD Pharmingen™ Alexa Fluor® 647 Mouse anti-SSEA-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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The MC813-70 monoclonal antibody reacts with Stage-Specific Embryonic Antigen-4 (SSEA-4), a carbohydrate epitope on the major ganglioside, but not the neutral glycolipid, of human teratocarcinoma cells. As its name implies, the expression of SSEA-4 is stage-specific and can be used to characterize embryonic cells and monitor their differentiation. However, its expression pattern differs in the human and mouse. In the human, SSEA-4 is found on teratocarcinoma (embryonal carcinoma or EC), embryonic inner cell mass (ICM), embryonic stem (ES) cells, and the K562 erythromyeloid leukeumia cell line. As human stem cells undergo differentiation, SSEA-4 expression is lost. In the mouse, SSEA-4 is found on oocytes and early cleavage-stage embryos, and primitive ectoderm, but not on EC, ICM, or ES cells. In some cases, SSEA-4 expression appears upon differentiation of mouse EC or ES cells.
Development References (7)
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Andrews PW, Bronson DL, Benham F, Strickland S, Knowles BB. A comparative study of eight cell lines derived from human testicular teratocarcinoma. Int J Cancer. 1980; 26(3):269-280. (Clone-specific). View Reference
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Draper JS, Pigott C, Thomson JA, Andrews PW. Surface antigens of human embryonic stem cells: changes upon differentiation in culture. J Anat. 2002; 200:249-258. (Clone-specific: Flow cytometry). View Reference
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Henderson JK, Draper JS, Baillie HS, et al. Preimplantation human embryos and embryonic stem cells show comparable expression of stage-specific embryonic antigens. Stem Cells. 2002; 20:329-337. (Clone-specific: Flow cytometry, Immunocytochemistry (cytospins)). View Reference
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Josephson R, Ording CJ, Liu Y, et al. Qualification of embryonal carcinoma 2102Ep as a reference for human embryonic stem cell research. Stem Cells. 2007; 25:437-446. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
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Kannagi R, Cochran NA, Ishigami F, et al. Stage-specific embryonic antigens (SSEA-3 and -4) are epitopes of a unique globo-series ganglioside isolated from human teratocarcinoma cells. EMBO J. 1983; 2(12):2355-2361. (Immunogen: Immunofluorescence, Radioimmunoassay). View Reference
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Son YS, Park JH, Kang YK, et al. Heat shock 70-kDa protein 8 isoform 1 is expressed on the surface of human embryonic stem cells and downregulated upon differentiation. Stem Cells. 2005; 23:1502-1513. (Clone-specific: Flow cytometry, Immunocytochemistry (cytospins)). View Reference
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Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282:1145-1147. (Clone-specific: Immunocytochemistry (cytospins)). 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.