Hpr conjugated secondary antibody

Proteins were resolved by SDS-PAGE, and transferred to nitrocellulose membranes for 1 h. Then, membranes were blocked 1 h at room temperature with TBS (tris-buffered saline), 5% BSA, and blotted overnight in TBS 5% BSA containing primary antibodies at 1:500 overnight with corresponding antibodies at 4 °C. Subsequently, the blot was washed and incubated with HPR-conjugated secondary antibody (Boster, Pleasanton CA, USA) for 1 h at room temperature at 1:5000 before being revealed with ECL (enhanced chemo-luminescence). The acquisition was performed with a Fusion FX7 imager (Vilber-Lourmat, Sud Torcy, France). The following primary antibodies were used: rabbit polyclonal anti-PARP (#9542, Cell Signalling, Danvers MA, USA), mouse polyclonal anti-CHOP (#2895, Cell Signalling), rabbit polyclonal anti-Phospho-IRE1α (#PA1-16927, Thermofisher Scientific), rabbit polyclonal anti-Phospho-eIF2α (#3398, Cell Signalling), and mouse monoclonal β-actin (#A5316, Sigma).

Protein extracts were resolved by SDS-PAGE and then transferred onto the nitrocellulose membranes for 1 h. Then, membranes were blocked for 1 h at room temperature with TBST (tris-buffered saline), 5% BSA, and blotted overnight in TBST 5% BSA containing primary antibodies at 1:500 overnight with corresponding antibodies at 4 °C. Subsequently, the blot was washed and incubated with HPR-conjugated secondary antibody (Boster, Pleasanton CA, USA) for 1 h at room temperature at 1:5000 before being revealed with ECL (enhanced chemo-luminescence). The acquisition was performed by a Fusion FX7 imagine system (Vilber-Lourmat, Sud Torcy, France). The following primary antibodies were used: rabbit polyclonal TFAM (# 7495, cell signaling), mouse monoclonal β-actin (#A5316, Sigma).