Mouse anti human cx43 primary antibodies

The control and cardiomyogenic induced groups were cultured on coverslips (Thermo scientific, UK) for 21 days. After fixation for 30 min at 4 °C with 4% paraformaldehyde, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at 4 °C, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 4 °C. After being washed with PBS, the cells were incubated with goat anti-mouse horseradish peroxidase secondary antibody (Immuno Tools GmbH, Germany) for 1 h at 37 °C. Finally, the immunoreaction was detected by using 3, 3 -diaminobenzidine substrate (Sigma-aldrich, USA). The cells were visualized under DMi1 inverted phase contrast microscope. The signal expression was analyzed using imageJ 1.50i software and calculated by CTCF = Integrated Density – (Area of selected cell * Mean signal of background readings). The data were presented as the mean ± SEM.

The control and cardiomyogenic induced groups were cultured on coverslips (Thermo scientific, UK) for 21 days. After fixation, the cells were blocked in 10% AB-serum in 1% BSA-PBS for 30 min at room temperature, and then incubated with mouse anti-human Cx43 primary antibodies (Sigma-aldrich, USA) for 2 h at 37 °C. After being washed with PBS, the cells were incubated with goat anti-mouse horseradish peroxidase secondary antibody (Immuno Tools GmbH, Germany) for 1 h at 37 °C. Finally, the immunoreaction was detected by using 3, 3 -diaminobenzidine (DAB) substrate (Sigma-aldrich, USA). The cells were visualized under Axiostar plus light microscope (Carl Zeiss, Germany). Photographs were taken with Canon pc 1049 PowershotG5 (Canon, USA). The signal expression was analyzed using imageJ 1.50i software and calculated by CTCF. $\text{CTCF}=\text{Integrated Density}-\left(\text{Area of selected cell}\ast \text{Mean signal of background readings}\right)$