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To ensure safety, pregnancy and breastfeeding are considered absolute GHK-Cu contraindications

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The overnight culture (2030 ml) was added to 1,000 ml of 2YT medium (containing 100 g/ml carbenicillin and 50 g/ml kanamycin) up to an optical density at 600 nm (OD 600 ) = 0.1 and grown at 37C until OD 600 = 0.6 before induction with 1 mM isopropyl--D-1- thiogalactopyranoside