Rabbit pan actin

Complete RIPA buffer was prepared by adding 1× Protease and Phosphatase inhibitor Cocktail (Thermo Scientific, Dreieich, Germany) with Pierce RIPA buffer (Thermo Scientific). Complete RIPA buffer were directly added to the cell culture plates or snap frozen tissues and then homogenized to isolate protein. The protein was denatured using 4× LDS sample buffer (Thermo Scientific) and 10× Reducing agent (Thermo Scientific). NuPAGE protein gels (Invitrogen, Carlsbad, CA, USA) were used to run the protein and PVDF membrane (Thermo Scientific) was used to transfer the protein from gel. Membranes were incubated with primary antibodies for overnight at 4 °C and with secondary antibodies for 1 h at RT. ECL substrate (GE Healthcare, Chicago, IL, USA) were added directly into the membrane for 2–3 min before capturing image at Amersham Imager 680 (GE Healthcare). Band intensities were also quantified using the software provided with the Imager.
Primary antibodies—Rabbit IRAG1, Rabbit PKG1α, Rabbit PKG1β (kindly provided by Prof. Schlossmann) [15 (link),25 (link)], Rabbit IP₃RI, Rabbit Pan-Actin (Cell Signaling Technology), Rabbit Serca2a, Rabbit pPLN-Thr 17, Mouse PLN (Badrilla, Leeds, UK).
Secondary antibodies—goat anti-rabbit, goat anti-mouse (Thermo Scientific).