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V450 Mouse anti-SSEA-4
V450 Mouse anti-SSEA-4
Flow cytometric analysis of BD Horizon™ V450 anti-SSEA-4 on human embryonic stem cells. H9 human embryonic stem (ES) cells (WiCell, Madison, WI) passage 37 grown grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested and stained with BD Horizon™ V450 anti-SSEA-4 (Cat. No. 561156, solid line histogram) or a BD Horizon™ V450 mouse IgG3, κ isotype control (Cat. No.561600, dashed line histogram). Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of BD Horizon™ V450 anti-SSEA-4 on human embryonic stem cells. H9 human embryonic stem (ES) cells (WiCell, Madison, WI) passage 37 grown grown in mTESR™1 media (StemCell Technologies) on BD Matrigel™ hESC-qualified Matrix (Cat. No. 354277) were harvested and stained with BD Horizon™ V450 anti-SSEA-4 (Cat. No. 561156, solid line histogram) or a BD Horizon™ V450 mouse IgG3, κ isotype control (Cat. No.561600, dashed line histogram). Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
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BD Horizon™
Stage-Specific Embryonic Antigen-4
Human (QC Testing), Mouse (Reported)
Mouse BALB/c IgG3, κ
Human Teratocarcinoma Cell Line
Flow cytometry (Routinely Tested)
5 µl
AB_10896140
Aqueous buffered solution containing protein stabilizer, glycerol 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™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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. 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.
  4. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. mTESR™1 is a trademark of StemCell Technologies.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561156 Rev. 3
Antibody Details
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MC813-70

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.

The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

561156 Rev. 3
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V450
Violet 405 nm
405 nm
450 nm
561156 Rev.3
Citations & References
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Development References (7)

  1. 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
  2. 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
  3. 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
  4. 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). View Reference
  5. 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). View Reference
  6. 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). View Reference
  7. 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
View All (7) View Less
561156 Rev. 3

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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.