The transformed Pichia pastoris cells are induced to express these proteins by adding 0.5% methanol daily to the culture, respectively
The structural difference is a single amino acid substitution at the first position: GHK-Cu uses glycine (Gly-His-Lys:Cu), while AHK-Cu uses alanine (Ala-His-Lys:Cu)
K., Mohammed, E., & Sant Bakshsingh, V
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Step 5: Inject
Over-lap PCR was used to generate six GSTP phosphorylation-mimicking mutants ( Co-Immunoprecipitation and Immunoblotting Analysis Cells were washed twice with phosphate-buffered saline (PBS), and then lysed in a lysis buffer on ice for 30 min, and then cell lysates were centrifuged at 12,500 rpm for 15 min