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I'm working on if a drug can enhance the ubiquitination of my protein of interest and thus increase its degradation. I've seen that the drug can downregulate the protein at 48hr but not at 24hr (or not quite obviously). Now I'm trying MG132 together with the drug to see if MG132 can block the drug's downregulation of the protein.
This is my first time using this drug, some suggest pre-treatment of MG132 before adding my drug, some suggest treatment of MG132 6hr pre-harvest. I'm not sure which one is better, but I am afraid if I pretreat and incubate with my drug for another 48hr, MG132 will be too toxic.
Can anyone give me any suggestions? THanks ahr!
