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For cell culture applications, prepare fresh dilutions into serum-free media immediately before use, as serum albumin and transferrin in complete media can sequester the copper ion over extended incubation periods, altering the effective GHK-Cu concentration

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The supernatant was incubated in the presence of 2 mM monobromobimane for 20 min at room temperature, and then acetic acid was added to the final concentration of 1% and incubated for 15 min on ice