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Also, if re-naturing the protein from inclusion bodies, be sure that all denaturants have been removed from the buffer, either through exhaustive dialysis or application of the sample to a desalting column, before applying to the glutathione column

In experiments in which large amounts of acetyl-CoA were incubated with purified enzyme to test its protection against SNO-CoA-mediated inhibition, varying levels of acetyl-CoA were added back into the assay solutions after the incubation period, such that in the assay condition, the concentration of acetyl-CoA was even across samples to control for the possible effect of acetyl-CoA as a competitive inhibitor of PDHC

In: Nature Reviews Neuroscience (2015)

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