Rabbit polyclonal anti oct4

The following antibodies were purchased from Abcam (Cambridge, MA): rabbit polyclonal anti-Oct4, anti-bax rabbit monoclonal, rabbit polyclonal anti-Sox2, rabbit polyclonal to Cyclin D1, mouse monoclonal IgG to β-actin, mouse anti-β-actin mAb, anti-Bcl-2 antibody, FITC- polyclonal goat anti-rabbit IgG. Rabbit anti-goat IgG—Rhodamine conjugate from Millipore (Billerica, MA); 4'-6′-diamidino-2-phenylindole nuclear stain (DAPI) and green phalloidin were purchased from Invitrogen (Carlsbad, CA). Anti-human CD44-PE, and anti-human CD24-FITC were purchased from BD Biosciences (Franklin Lakes, NJ). PE-anti-rabbit IgG was purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Mouse iPSCs (SC201A-iPSC, passage 3, System Biosciences, Mountain View, CA, USA) were maintained in ESGRO complete plus clonal grade medium. The cells were passaged on reaching 80%–90% confluence at a ratio of 1:4 in 75 cm² flasks pre-coated with 0.1% gelatin. iPSCs of passages 11–13 were used for all experiments. The iPSCs were checked for SSEA-1, SOX2, Nanog and Oct4 expression by immunocytochemistry with the respective antibodies: mouse monoclonal anti-SSEA1-PE antibody (1:100, Millipore, Temecula, CA, USA), rabbit polyclonal anti-SOX2 (1:1000, Abcam, Cambridge, MA, USA), rabbit polyclonal anti-Nanog (1:700, Abcam) and rabbit polyclonal anti-Oct4 (1:1000, Abcam) at 4 °C overnight and fluorescein isothiocyanate-conjugated secondary antibody (1:100, Millipore) at room temperature for 1 h, images were captured by a fluorescence microscope (Eclipse E600, Nikon, Tokyo, Japan).