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Then the membrane was blocked with 5% BSA for 1 h at room temperature, followed by incubating with primary antibodies and IRDye800 flurophore-conjugated antibodies (LI-COR Biosciences, Lincoln, NE, USA)

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The process itself is quite structured

The PVDF membranes were incubated with the appropriate primary antibodies overnight at 4 C with gentle shaking, and then incubated with a goat anti-mouse/rabbit IgG horseradish peroxidase-conjugated antibody

current methods of hydrogen delivery, including inhalation and hydrogen-enriched water, exhibit significant variability in hydrogen concentration within brain tissues