Western Blotting of Mitochondrial Proteins

Western blotting was performed as previously described (Niemi et al., 2019 (link)). Antibodies used in this study include: anti-Pptc7 (Novus Biologicals, catalog #NBP1-90654, 1:1000 dilution, 48 hour incubation), anti-Citrate synthase (Cell Signaling Technologies, catalog #14309, 1:1000 dilution, overnight incubation), anti-beta actin (Abcam, catalog # ab170325, 1:1000 dilution, overnight incubation), anti-Bnip3 (rodent specific antibody, Cell Signaling Technologies, catalog #3769, 1:1000 dilution, 48 hour incubation), and anti-Nix (Cell Signaling Technologies, catalog #12396, 1:1000 dilution, overnight incubation).

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Niemi N.M., Serrano L.R., Muehlbauer L.K., Balnis C., Kozul K.L., Rashan E.H., Shishkova E., Schueler K.L., Keller M.P., Attie A.D., Pagan J., Coon J.J, & Pagliarini D.J. (2023). Pptc7 maintains mitochondrial protein content by suppressing receptor-mediated mitophagy. bioRxiv.

Publication Preprint 2023

NBP1-90654 anti-Citrate synthase

Antibodies Antibody Beta actin Biologicals Citrate synthase Dilution Novus Rodent

Corresponding Organization : University of Wisconsin–Madison

Other organizations : University of Queensland, Quantitative BioSciences

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Variable analysis

independent variables

Antibodies used in this study
Antibody dilutions
Antibody incubation times

dependent variables

Protein expression levels

control variables

Western blotting protocol (previously described in Niemi et al., 2019)

controls

Positive control: Not specified
Negative control: Not specified

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