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562706_IMAGE1.png

Flow cytometric analysis of SSEA-3 expression of human embryonic stem (ES) cells.  H9 human ES cells (WiCell, Madison, WI) passage 41 grown in mTESR™1 media (StemCell Technologies) were harvested with Accutase™ (Cat. No. 561527) and stained with BD Horizon™ BV421 Rat Anti-SSEA-3 (solid line) or a BD Horizon™ BV421 rat IgM, κ isotype control (Clone R4-22, Cat. No.562708, dashed line). Flow cytometry was performed on a BD™ LSRII flow cytometry system.

562706_IMAGE1.png Blue-Cap-Violet-Label.png

Flow cytometric analysis of SSEA-3 expression of human embryonic stem (ES) cells.  H9 human ES cells (WiCell, Madison, WI) passage 41 grown in mTESR™1 media (StemCell Technologies) were harvested with Accutase™ (Cat. No. 561527) and stained with BD Horizon™ BV421 Rat Anti-SSEA-3 (solid line) or a BD Horizon™ BV421 rat IgM, κ isotype control (Clone R4-22, Cat. No.562708, dashed line). Flow cytometry was performed on a BD™ LSRII flow cytometry system.

Product Details
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BD Horizon™
Human (QC Testing), Mouse (Reported)
Rat F344, also known as Fischer, CDF IgM
Mouse Embryos
Flow cytometry (Routinely Tested)
5 µl
AB_2737734
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
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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. Brilliant Violet™ 421 is a trademark of Sirigen.
  4. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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562706 Rev. 2
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.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562706 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
562706 Rev.2
Citations & References
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View product citations for antibody "562706" on CiteAb

Development References (5)

  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). 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). 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). 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). View Reference
  5. Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282:1145-1147. (Clone-specific). View Reference
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562706 Rev. 2

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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.