Goat anti human oct4

Immunofluorescence staining of the riPSC90 cell line was performed as
previously published (Sosa et al., 2016 (link)).
In short, riPSC90 colonies were fixed in 4% PFA, permeabilized with PBS
plus 0.5% Triton™ X-100 (Sigma) then washed in PBST (PBS with
0.1% Tween-20 (Sigma)) Primary antibodies included goat-anti-human OCT4
(1:100, Santa Cruz), goat-anti-human NANOG (1:100, R&D Systems),
mouse-anti-human SSEA4 (1:100, Developmental Studies Hybridoma Bank), and
mouse-anti-human TRA-1-81 (1:100, eBiosciences). For detecting primary
antibodies raised in goat, AF594-conjugated donkey anti-goat (1:100, Life
Technologies) secondary antibodies were used. For detecting primary antibodies
raised in mice, AF488- or AF594-conjugated donkey-anti-mouse (1:100, Life
Technologies) secondary antibodies were used. Images of cells were acquired with
a Zeiss LSM 880 confocal laser-scanning microscope. Images were analysed using
ImageJ version 1.51d (NIH).

riPSC89 cells were fixed in 4% PFA, permeabilized with PBS plus 0.5% Triton™X-100 (Sigma) then washed in PBST (PBS with 0.1% Tween-20 (Sigma)) and blocked in 10% donkey serum (Jackson Immunoresearch) for 30 minutes at room temperature. After washing in PBST, cells were incubated overnight at 4°C with primary antibodies diluted in PBS containing 2.5% donkey serum. The primary antibody concentrations were, goat-antihuman OCT4 (1:100, Santa Cruz), goat-anti-human NANOG (1:100, R&D Systems), mouse-anti-human SSEA-4 (1:100, Developmental Studies Hybridoma Bank), and mouse-anti-human TRA-1-81 (1:100, eBiosciences). After washing in PBST, the cells were incubated for 1 hour at room temperature with secondary antibodies in PBS containing 2.5% donkey serum. For sections stained with primary antibodies raised in goat, AF594-conjugated donkey anti-goat (1:100, Life Technologies) was used. For detecting primary antibodies raised in mice, AF488- or AF594-conjugated donkey-anti-mouse (1:100, Life Technologies) were used. Cells were then washed in PBST prior to mounting under coverslips with Prolong Gold Antifade mountant with DAPI (Life Technologies). Images of cells were acquired with a Zeiss LSM 880 confocal laser-scanning microscope. Images were analysed using ImageJ version 1.51d (NIH).