Rabbit polyclonal anti oct4 antibody

Fixed samples were washed with PBS to remove paraformaldehyde, and then rabbit polyclonal anti–OCT4 antibody (1:1000) (Abcam, USA) in PBST (PBS+0.1% Triton-X) supplemented with 10% normal goat serum was added to each well and incubated overnight at 4°C. After rabbit polyclonal anti-OCT4 antibody was removed and samples were washed with PBS, goat anti-rabbit Alexa-Fluor 568 (1:500) in PBS-Triton supplemented with 10% NGS were added to each well and incubated for 45 min at RT in dark. DAPI (1:500) was also added to goat anti-rabbit Alexa-Fluor 568 solution to counterstain cell nuclei. About 10–15 characteristic pictures/well were taken by a fluorescent microscope (Olympus IX-71) equipped with a Canon EOS digital camera (Canon, Tokyo, Japan). Then, cells were counted after merging images by Adobe Photoshop CS4, and the results were displayed as a percentage of total cells counted.