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Alexa Fluor® 647 Rat Anti-SSEA-3
Alexa Fluor® 647 Rat Anti-SSEA-3

Flow cytometric analysis of SSEA-3 expression on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 37 grown on irradiated mouse embryonic fibroblasts were harvested and stained with either Alexa Fluor® 647 rat anti-SSEA-3 antibody (solid line) or an Alexa Fluor® 647 rat IgM, κ isotype control (Clone R4-22, Cat. No.560892, dashed line). Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Alexa Fluor® 647 Rat Anti-SSEA-3

Immunoflourescent staining of SSEA-3 on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 33 grown on irradiated mouse embryonic fibroblasts were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and stained with Alexa Fluor® 647 rat anti-SSEA-3 monoclonal antibody (pseudo-colored red) at 10 μg/mL. Cell nuclei were stained with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.

Flow cytometric analysis of SSEA-3 expression on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 37 grown on irradiated mouse embryonic fibroblasts were harvested and stained with either Alexa Fluor® 647 rat anti-SSEA-3 antibody (solid line) or an Alexa Fluor® 647 rat IgM, κ isotype control (Clone R4-22, Cat. No.560892, dashed line). Flow cytometry was performed on a BD™ LSR II flow cytometry system.

Immunoflourescent staining of SSEA-3 on human embryonic stem (ES) cells. H9 human ES cells (WiCell, Madison, WI) passage 33 grown on irradiated mouse embryonic fibroblasts were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and stained with Alexa Fluor® 647 rat anti-SSEA-3 monoclonal antibody (pseudo-colored red) at 10 μg/mL. Cell nuclei were stained with Hoechst 33342 (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software.

Product Details
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BD Pharmingen™
Human (QC Testing), Mouse (Reported)
Rat F344, also known as Fischer, CDF IgM
Mouse Embryos
Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
5 µl
AB_10893794
Aqueous buffered solution containing BSA, 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, see the protocol at http://www.bdbiosciences.com/support/resources/protocols/ceritifed_reagents.jsp.

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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. 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.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  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 refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561145 Rev. 1
Antibody Details
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MC-631

The MC-631 monoclonal antibody reacts with Stage-Specific Embryonic Antigen-3 (SSEA-3), a carbohydrate epitope on the major ganglioside, but not the neutral glycolipid, of mouse embryos and human teratocarcinoma cells.  As its name implies, the expression of SSEA-3 is stage-specific and can be used to characterize embryonic cells and monitor their differentiation.  However, its expression pattern differs between human and mouse.  In the human, SSEA-3 is found on teratocarcinoma (embryonal carcinoma or EC), embryonic inner cell mass (ICM), embryonic stem (ES) cells, and erythrocytes.  As human stem cells undergo differentiation, SSEA-3 expression diminishes.  In the mouse, SSEA-3 is found on oocytes, ova, zygotes, early cleavage-stage embryos, early blastocysts, ICM, primitive endoderm, and adult kidneys, but not on EC or ES cells.

561145 Rev. 1
Format Details
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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
561145 Rev.1
Citations & References
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Development References (6)

  1. 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
  2. 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: Cytotoxicity, Flow cytometry, Immunocytochemistry (cytospins)). View Reference
  3. 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. (Clone-specific: Immunofluorescence, Radioimmunoassay). View Reference
  4. Shevinsky LH, Knowles BB, Damjanov I, Solter D. Monoclonal antibody to murine embryos defines a stage-specific embryonic antigen expressed on mouse embryos and human teratocarcinoma cells. Cell. 1982; 30:697-705. (Immunogen: Cytotoxicity, Immunofluorescence, Immunoprecipitation, Radioimmunoassay). View Reference
  5. 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
  6. 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
View All (6) View Less
561145 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.

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.