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The cells were lysed for 5 min on ice in IP buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% NP-40, 5 mM EDTA, 5 mM EGTA) containing additionally 50 mM NaF, 1 mM vanadate, 1 mM PMSF, 10 g/mL TPCK and 1x protease inhibitor cocktail (Roche) and centrifuged for 10 min at 20,000 g and 4 C

Once or twice daily application is the typical protocol for copper peptide scalp serums

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