Rabbit anti vdac1 porin

Human, mouse and cellular protein extracts were homogenized in RIPA buffer and incubated in the following primary antibodies: mouse anti-Tim23 (1:1000, BD Biosciences), mouse anti-Tom20 (1:1000, Abcam), mouse anti-Total OXPHOS (1:1000, Abcam, ab110411), rabbit anti-eGFP (1:1000, Novus Biologicals, Littleton, CO, USA, NBP2-37821), rabbit anti-ClpX (1:2000, Abcam, ab168338), mouse anti-Grp75 (1:1000, Abcam, ab2799), mouse anti-Ndufs3 (1:1000, Abcam), rabbit anti-COX IV (1:1000, Abcam), rabbit anti-VDAC1/Porin (1:2000, Abcam, ab15895), mouse anti-α-Tubulin (1:5000, Sigma-Aldrich, T5168) and mouse anti-β-actin (1:5000, Sigma-Aldrich, A5441).

Western blots were performed as described previously (Suzuki et al., 2015 (link)). Briefly, mitochondrial fractions and cytosolic fractions were isolated using a mitochondria isolation kit for cultured cells (Thermo Scientific, Rockford, IL). Equal amounts of protein per lane (5–20 μg) were loaded onto Mini-Protean® TGX™ gels (Biorad, Hercules, CA), transferred to Trans-Blot Turbo Transfer nitrocellulose membranes (Biorad) and probed with primary antibodies. Primary antibodies included: rabbit anti-cytochrome-c, rabbit anti-VDAC1/Porin (Abcam, Inc., Cambridge, MA), and rabbit anti-β-actin (Cell Signaling Technology, Danvers, MA). The secondary antibody was HRP-conjugated goat anti-rabbit IgG (Biorad). Enhanced chemiluminescence was performed with SuperSignal West Pico (Thermo Scientific). Signal was detected by myECL imager (Thermo Scientific) and band density was quantified using myimageAnalysis™ Software (Thermo Scientific).