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We therefore sought a strategy to reduce the scale of the library to remain within the quantitative throughput limits of our assay (up to ~40,000 substrates), while still assaying a full repertoire of sequences likely to be cleaved by our evolved proteases (Extended Data Fig
Western blot analysis involved preparing whole cell lysates using RIPA lysis buffer, quantifying protein concentration via BCA assay, followed by SDS-PAGE, transfer to nitrocellulose membranes, incubation with primary and HRP-conjugated secondary antibodies, and detection using an ECL solution [1]
This kind of trial is expensive typically several million dollars and to date has been more difficult to justify economically because GHK-Cu cannot be patented as a novel drug (the molecule is endogenous and the relevant patents have expired)
A dosage of 2ml is recommended as safe, anything more could cause irritation at the injection site
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