Goat anti sox17 antibody

For whole-mount immunocytochemical analysis, iPS cells cultures were fixed in 4 % paraformaldehyde (FUJIFILM Wako Pure Chemical Corporation) for 30 min, followed by permeabilization with 0.1 % Triton-X (nakalai tesque) in phosphate-buffered saline (PBS; Thermo Fisher Scientific) for 10 min at room temperature, washed once with 0.1 % Tween-20 (MP, Irvine, CA, USA) in PBS (PBST) then incubated with 20 % Blocking One (nakalai tesque) in PBS-T in a humidified chamber for 1 h at room temperature. The cells were incubated with diluted (1:200) antibody in 20 % Blocking One in PBS-T. After washing 3 times with PBS-T, the cells incubated with diluted (1:1000) secondary antibody in 20 % Blocking One for 2 h at room temperature in the dark. After washing off the secondary antibody in PBS-T, cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). The following antibodies were used as first antibodies: Mouse anti-OCT3/4 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), goat anti-SOX17 antibody (R&D systems), Goat anti-human Albumin antibody (Fortis Life Sciences, Montgomery, TX, USA). Secondary antibodies used were Alexa 488-conjugated and Alexa 568-conjugated antibodies (Thermo Fisher Scientific).