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PA homogenates (200 µg) were used to immunoprecipitate RyR2 with anti-RyR2 antibody (Thermo Scientific). Then add the protein-G (Santa Cruz Biotech) and incubate at 4 °C overnight. The immunoprecipitates were washed and then elute by adding Laemmli sample buffer. After the samples were boiled for 5 min, equal homogenate was separated by SDS-PAGE (6% gel for RyR2, 15% gel for FKBP12.6). Proteins were transferred to PVDF membrane and develop using anti-RyR2 antibody (1:1000), anti-FKBP12.6 antibody (1:800), then visualized by the chemiluminescence detection kit13.
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