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HUBERMAN: And this is very important to understand because unlike a lot of our discussion on this podcast about dopamine or about specific hormones like testosterone or estrogen, when we talk about peptides, especially therapeutic peptides, oftentimes we're talking about a single peptide that does a bunch of different things depending on the cell type, the time of day or night that that peptide is present, and even the different so-called downstream effects of the peptide

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Harvested cells were resuspended in 20 mM HEPES pH 7.0, 500 mM NaCl, 25 mM imidazole (lysis buffer) supplemented with protease inhibitor cocktail and then lysed by a cell disruptor at 25 kpsi (Constant Systems)