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Next, membranes were blocked with 5% BSA in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) for 1 h at room temperature and probed with primary antibodies against corresponding antigens overnight at 4 C

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You get a more sustained, non-physiologic elevation

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It also interacts with neurotransmitter systems, supporting mood balance, stress resilience, and potential neuroprotective effects