Mouse monoclonal anti sox2

iPSCs were cultured on matrigel-coated coverslips, fixed in paraformaldehyde and permeabilized in blocking/permeabilization buffer (2% BSA/2% FBS/0.05% NP-40 in PBS) for 45 min and incubated with primary antibodies overnight at 4°C. Then the cells were washed in PBS and incubated with Alexa-conjugated secondary antibodies (Invitrogen) diluted in blocking/permeabilization buffer (1:750). Finally, after washing in PBS the cells were counterstained with DAPI. Immunofluorescence images were acquired using an Olympus X41 microscope. The following antibodies were used: mouse monoclonal anti-OCT4 (Santa Cruz biotechnology, Germany), goat polyclonal anti-NANOG (R and D systems), mouse monoclonal anti-SOX2 (R and D systems), mouse monoclonal anti-SSEA-4 (R and D systems, Minneapolis, MN), mouse monoclonal anti-TRA-1–60 (R and D systems). Similarly, iPSC-derived cardiomyocytes were dissociated and cultured on matrigel-coated coverslips for 4–5 days, fixed in paraformaldehyde and permeabilized in blocking/permeabilization buffer for 45 min. The following primary antibodies were used: mouse monoclonal anti-cardiac troponin T (Thermo Fisher Scientific, France), mouse monoclonal anti-connexin 43, and mouse monoclonal anti-α-actinin. Confocal imaging was performed using a Leica SP5 confocal system.

iPSC were cultured on matrigel-coated coverslips, fixed in paraformaldehyde and permeabilized in blocking/permeabilization buffer (2% BSA/2% FBS/0.05% NP40 in PBS) for 45 min and incubated with primary antibodies overnight at 4 °C. Then the cells were washed in PBS and incubated with Alexa-conjugated secondary antibodies (Invitrogen) diluted in blocking/permeabilization buffer (1:750). Finally, after washing in PBS the cells were counterstained with 4,6-diamidino-2-phenylindole (DAPI). Immunofluorescence images were acquired using an Olympus X41 microscope. The following antibodies were used: mouse monoclonal anti-OCT4 (1:100, Santa Cruz), goat polyclonal anti-NANOG (1:100, R&D systems), mouse monoclonal anti-SOX2 (1:100, R&D systems) and mouse monoclonal anti-SSEA-4 (1:100, R&D systems). Similarly, iPSC-derived CMs were dissociated and cultured on matrigel-coated coverslips for 4–5 days, fixed in paraformaldehyde and permeabilized in blocking/permeabilization buffer for 45 min. The cells were incubated with Alexa-conjugated primary antibodies overnight at 4 °C, washed in PBS and counterstained with DAPI. Confocal imaging was performed using a Leica SP5 confocal system. The following antibodies were used: mouse monoclonal anti-cardiac troponin T (1:200, Thermo Fisher Scientific) and mouse monoclonal anti-PLN (1:200, Bradilla).