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Biotinylated proteins detected by blotting with streptavidin-HRP. (Lane 1) Unmodified histones were labeled with biotin at free lysines and N termini. (Lanes 2 and 3) Acetylated histones were not labeled because they did not have any free amines. (Lane 4) Acetylated histones in the presence of sirtuin and NAD+ were deacetylated, and these free lysines were biotinylated, allowing them to be detected by the streptavidin-HRP.
