BN-PAGE was performed as described previously (Van Vranken et al., 2018 (link)). 100 μg of mitochondria were resuspended in 1× lysis buffer (Invitrogen, BN20032) supplemented with yeast protease inhibitor cocktail (Sigma, P8215) and solubilized with 1% digitonin for 20 min on ice. Solubilized mitochondria were cleared by centrifugation at 20,000 × g for 20 min. Lysate was mixed with NativePAGE 5% G-250 Sample Additive (Invitrogen, BN20041) and resolved on a 3–12% gradient native gel (Invitrogen, BN1001BOX).
Other organizations :
University of Utah, Harvard University, Koç University, University of Colorado Boulder, University of Helsinki, Czech Academy of Sciences, Institute of Biotechnology, Howard Hughes Medical Institute
Mitochondrial protein complexes resolved on the native gel
control variables
Lysis buffer used (1× lysis buffer from Invitrogen, BN20032)
Protease inhibitor cocktail used (yeast protease inhibitor cocktail from Sigma, P8215)
Native gel used (3–12% gradient native gel from Invitrogen, BN1001BOX)
Sample preparation (Lysate mixed with NativePAGE 5% G-250 Sample Additive from Invitrogen, BN20041)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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