Rabbit anti hsp60 antibody

Protein extracts from wt and CG1603 mutant larvae tissues (48h after egg laying) were prepared using the RIPA buffer (MilliporeSigma) with Halt Protease Inhibitor Cocktail (Thermo Fisher Scientific), 5 mM NaF (MilliporeSigma) and 1 mM Na3VO4 (MilliporeSigma). Western blot was performed using a XCell SureLock^™ Mini-Cell and XCell II^™ Blot Module (Thermo Fisher Scientific). Samples were electrophoresed under a reducing condition on NuPAGE^™ 4 to 12% Bis-Tris Mini Protein Gels (Thermo Fisher Scientific). Proteins on the gel were transferred to a Polyvinylidene Difluoride (PVDF) membrane (Thermo Fisher Scientific). The membranes were blocked with 5% BSA or non-fat milk (MilliporeSigma) in TBST (Tris buffered saline with 0.1% Tween-20). After a serial of washes and incubations with primary antibodies, TBST and secondary antibodies, the immunoreactivity was visualized using SuperSignal West Dura Chemiluminescent Substrate (Thermo Fisher Scientific) and Amersham ImageQuant 800 system (Cytiva). The antibodies used were: Mouse anti-Actin antibody (C4, MilliporeSigma), Mouse anti-ATP5A antibody (15H4C4, abcam), Mouse anti-ND30 antibody (17D95, abcam), rabbit anti-TFAM antibody (Liu et al., 2022 (link)), rabbit anti-HSP60 antibody (#4870, Cell Signaling), Anti-rabbit IgG, HRP-linked Antibody (#7074, Cell Signaling) and Anti-mouse IgG, HRP-linked Antibody (#7076, Cell Signaling).