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Flow cytometric analysis of SSEA-1 expression on E14 cells. E14 mouse embryonic stem (ES) cells were stained with either PE Mouse Anti-SSEA-1 (Cat. No. 560142/560886; solid line histogram) or PE Mouse IgM, κ Isotype Control (Cat. No. 555584, dashed line histogram), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Fluorescence histogram depicting SSEA-1 (or Ig isotype expression) were derived from gated events with the side and forward light-scattering characteristics of viable cells. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of SSEA-1 expression on E14 cells. E14 mouse embryonic stem (ES) cells were stained with either PE Mouse Anti-SSEA-1 (Cat. No. 560142/560886; solid line histogram) or PE Mouse IgM, κ Isotype Control (Cat. No. 555584, dashed line histogram), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Fluorescence histogram depicting SSEA-1 (or Ig isotype expression) were derived from gated events with the side and forward light-scattering characteristics of viable cells. Flow cytometry was performed on a BD FACSCalibur™ System.
Flow cytometric analysis of SSEA-1 expression on E14 cells. E14 mouse embryonic stem (ES) cells were stained with either PE Mouse Anti-SSEA-1 (Cat. No. 560142/560886; solid line histogram) or PE Mouse IgM, κ Isotype Control (Cat. No. 555584, dashed line histogram), incubated in the dark for 20 minutes at room temperature and analyzed by flow cytometry. Fluorescence histogram depicting SSEA-1 (or Ig isotype expression) were derived from gated events with the side and forward light-scattering characteristics of viable cells. Flow cytometry was performed on a BD FACSCalibur™ System.
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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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
The MC480 monoclonal antibody reacts with Stage-Specific Embryonic Antigen-1 (SSEA-1), which is a terminal carbohydrate epitope (3-fucosyl-N-acetyllactosamine or 3-FAL) on glycoproteins and lactose-series glycolipids. SSEA-1 is related to Lewis blood group antigens and is found in a variety of embryonic as well as adult tissues and cancers. As its name implies, the expression of SSEA-1 is stage-specific and can be used to characterize embryonic cells and monitor their differentiation. However, its expression pattern differs between human and mice. In the human, SSEA-1 is not found on embryonic stem (ES) cells, embryonic inner cell mass (ICM), or teratocarcinoma (embryonal carcinoma or EC) cells. As human EC and ES cells undergo differentiation, SSEA-1 expression is upregulated. In the adult, the same epitope is expressed as CD15 on granulocytes and monocytes, but not lymphocytes or dendritic cells. In the mouse, SSEA-1 is found on EC, ES, primordial germ cells, 8-cell to blastocyst embryos, ICM, and subpopulations of cells in the adult central nervous system, including stem cells. In contrast to human SSEA-1 expression, it is reduced when mouse EC and ES cells undergo differentiation.
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