Human induced pluripotent stem cells (iPSCs) stained with ab16287 (red fill) with secondary only control (blue fill). In brief, iPSCs were fixed in 4% formaldehyde (methanol-free) for 15 min at 25°C. Cells were then incubated with the antibody (ab16287, 1/9600 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG at 1/1000 dilution for 30 min at 4°C. iPSCs differentiated for 14 days toward a cardiomyocyte lineage were used as a negative control. Acquisition of >20,000 total events were collected using a 100mW solid state diode laser (488nm) and 529/28 bandpass filter.

ab16287 staining SSEA4 in rat tendon derived stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 in PBS and blocked with 1% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody (1/100) for 20 hours at 4°C. An Alexa Fluor® 594-conjugated donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

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Immunofluorescence analysis of Human dental pulp stem cells, staining SSEA4 with ab16287 at 1/40 dilution. A FITC-conjugated anti-mouse IgG was used as the secondary antibody.

Immunocytochemistry/ Immunofluorescence analysis of human iPSC cells labeling SSEA4 with ab16287 at 1/500 dilution. Cells were fixed with paraformaldehyde and  permeabilized with 0.5% TX100. Cells were blocked with 5% serum for 20 minutes at 25°C. A goat polyclonal anti-mouse Cy3 secondary antibody at 1/500 dilution was used. 

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anti-SSEA4 antibody, ab16297 can be used as a marker of Embryonic Carcinoma cells in Flow Cytometry/FACS. As can be seen from the histograms, in non-differentiating conditions (i.e. without retinoic acid) NTERA2 cells were recognised by ab16297. However, upon differentiation (addition of retinoic acid), the antibody lost the ability to recognise the cells.

2102ep Human Embryonic Carcinoma cells were stained with SSEA4 antibody ab16287. As expected, staining localised to the cell surface (green). Nuclei are stained blue using Hoechst.

ab16287 staining SSEA4 in human Embryonic Stem Cells, HUES7 by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with Triton and blocking with 10% serum for 1 hour was performed. Samples were incubated with primary antibody (1/100: in 1% serum, 0.1% Triton in PBS) for 1 hour at 37°C. An Alexa Fluor^® 588-conjugated goat polyclonal to mouse IgG was used at dilution at 1/100 as secondary antibody.

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