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doi: 10.1007/s12031-019-1257-9 170 RushA
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The nominal concentration and the free concentration are different quantities, and a serum-free comparison is not measuring the same exposure
Because copper itself has a narrow physiological range, protocols involving repeated or high-frequency application in animal models often include copper-load monitoring alongside the primary remodeling endpoints, to rule out confounding from excess trace-mineral exposure rather than peptide-specific signaling
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