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Investigating Toxic Overload and Depression A targeted history and laboratory assessment may help distinguish environmental exposure from elevated SAH, glutathione depletion, mitochondrial stress, gut-derived burden or impaired organ clearance
The membranes were incubated with primary antibodies LC3A/B (1:1000 dilution, #4108, Cell Signaling Technology, Massachusetts, United States), p62 (1:2000 dilution, ab56416, Abcam, Massachusetts, United States), PINK1 (1:1000 dilution, 23274-1-AP, Proteintech Group, Illinois, United States), Parkin (1:1000 dilution, ab15954, Abcam, Massachusetts, United States), and GAPDH (1:2000 dilution, #2118, Cell Signaling Technology, Massachusetts, United States) overnight at 4C after blocking the membranes with 5% nonfat milk for 1 h, and then incubated with the appropriate IRDye-conjugated secondary antibody (1:10000 dilution, IRDye 800 CW goat anti-rabbit or IRDye 680RD goat anti-mouse, LI-COR Biosciences, Nebraska, United States) and imaged using the LI-COR Odyssey
Major challenges in this method include maintaining stability and compatibility of multiple agents, avoiding unwanted interaction, and optimizing pharmacokinetics and pharmacodynamics
Prochaska & Benowitz, 2016
