Hi,
I am planning to do Acyl Biotin exchange assay on red blood cell ghosts to detect the palmitoylome of the RBCs. Also, I want to treat the RBC ghosts with 2-BP and palimotoylation inhibitor and then determine the palmitoylome as well. For 2-BP treated RBCs, I expect to see a reduction in over all palmitoylation compared to untreated. I was wondering for such assays what should be my control? Typically following pull down of palmitoylated proteins, people run SDS-PAGE. If I see a decrease in protein content following 2-BP treatment, how could I convincingly say that there is a decrease in over all palmitoylation? Do I need to run total lysates both from control and 2-BP treated RBC ghosts, to show that the over all protein content was similar?
Thanks for your suggesitons
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