Western Blotting Protocol for Protein Analysis

Western blotting was performed following previous laboratory procedures [15 (link), 28 , 39 (link)]. The following primary antibodies were used for blotting: anti-parkin antibody (Abcam, catalog no. ab77924; Santa Cruz Biotechnology, catalog no. sc-32282), PINK1 antibody (Novus Biologicals, catalog no. BC100–494), caspase 3 polyclonal antibody (Proteintech, catalog no. 19677-1-AP), anti-optineurin antibody [EPR20654] (Abcam, catalog no. ab213556), phospho-TBK1/NAK (Ser172) (D52C2) XP rabbit monoclonal antibody (Cell Signaling Technology, catalog no. 5483), LC3 rabbit polyclonal antibody (Proteintech, catalog no. 14600-1-AP), TOMM40 rabbit polyclonal antibody (Proteintech, catalog no. 18409-1-AP), SOD2 rabbit polyclonal antibody (Proteintech, catalog no. 24127-1-AP), COXIV rabbit polyclonal antibody (Proteintech, catalog no. 11242-1-AP), GAPDH monoclonal antibody (Proteintech, catalog no. 60004-1-Ig), beta-actin monoclonal antibody (Proteintech, catalog no. 66009-1-Ig), and alpha-tubulin polyclonal antibody (Proteintech, catalog no. 11224-1-AP). Secondary antibodies included HP-goat anti-mouse (ZsBio, catalog no. ZB-2305Beijing, China) and HP-goat anti-rabbit (ZsBio, catalog no. ZB-2301). The blots were visualized using a 5200 Chemiluminescent Imaging System (Tanon Science and Technology).

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Li J., Lai M., Zhang X., Li Z., Yang D., Zhao M., Wang D., Sun Z., Ehsan S., Li W., Gao H., Zhao D, & Yang L. (2022). PINK1-parkin-mediated neuronal mitophagy deficiency in prion disease. Cell Death & Disease, 13(2), 162.

Publication 2022

PINK1 antibody caspase 3 polyclonal antibody LC3 rabbit polyclonal antibody GAPDH monoclonal antibody beta-actin monoclonal antibody alpha-tubulin polyclonal antibody

Alpha tubulin Anti antibody Antibodies Antibody Beta actin Biologicals Caspase 3 Gapdh Goat Monoclonal antibody Mouse Novus Optineurin Parkin Rabbit Sod2

Corresponding Organization :

Other organizations : China Agricultural University

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Variable analysis

independent variables

Primary antibodies used for blotting: anti-parkin antibody (Abcam, catalog no. ab77924; Santa Cruz Biotechnology, catalog no. sc-32282), PINK1 antibody (Novus Biologicals, catalog no. BC100–494), caspase 3 polyclonal antibody (Proteintech, catalog no. 19677-1-AP), anti-optineurin antibody [EPR20654] (Abcam, catalog no. ab213556), phospho-TBK1/NAK (Ser172) (D52C2) XP rabbit monoclonal antibody (Cell Signaling Technology, catalog no. 5483), LC3 rabbit polyclonal antibody (Proteintech, catalog no. 14600-1-AP), TOMM40 rabbit polyclonal antibody (Proteintech, catalog no. 18409-1-AP), SOD2 rabbit polyclonal antibody (Proteintech, catalog no. 24127-1-AP), COXIV rabbit polyclonal antibody (Proteintech, catalog no. 11242-1-AP), GAPDH monoclonal antibody (Proteintech, catalog no. 60004-1-Ig), beta-actin monoclonal antibody (Proteintech, catalog no. 66009-1-Ig), and alpha-tubulin polyclonal antibody (Proteintech, catalog no. 11224-1-AP)

dependent variables

Not explicitly mentioned

control variables

Western blotting was performed following previous laboratory procedures [15 (link), 28 (no link found), 39 (link)]
Secondary antibodies included HP-goat anti-mouse (ZsBio, catalog no. ZB-2305Beijing, China) and HP-goat anti-rabbit (ZsBio, catalog no. ZB-2301)
The blots were visualized using a 5200 Chemiluminescent Imaging System (Tanon Science and Technology)

controls

No positive or negative controls explicitly mentioned

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