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Membranes were stripped for 20 min at room temperature using a Blot Restore Membrane Rejuvenation kit (Sigma-Aldrich, 2520-M), then washed twice with TBS-T, re-blocked with BSA 5% for 30 min and incubated with mouse-anti--actin (Sigma-Aldrich, A1978) 1:3,000 in BSA 2.5% for 4 h at room temperature and then incubated with a sheep-anti-mouse (Amersham, NA931) secondary antibody at 1:5,000 in BSA 2.5% for 2 h

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Users combining 5-Amino-1MQ with exogenous ketones or MCT oil report enhanced subjective energy during fasted training, likely due to synergistic NAD+ elevation and mitochondrial substrate availability