Figure 5. Immunodetection of Cardiac Lysine Acetylation.

Panel A. Guinea pig cardiac myofilaments were prepared essentially as described by Solaro et al. [70]. 30 µg of protein was subjected to electrophoresis and immunoblotting with both monoclonal and polyclonal antibodies to acetylated lysine as described in the Methods section. Lanes with immunoreactive bands are juxtaposed with the corresponding exposure of a parallel control experiment conducted with >2% w/v acetylated BSA in the primary antibody incubation step. Panels B and C. Macromolecular respiratory complexes were resolved as described in the methods. Up to 20 µg of protein from the 30% w/v sucrose fraction (enriched Complex I, Panel B) and the 22.5% w/v sucrose fraction (enriched complex V, Panel C) were probed for immunoreactivity to anti-acetylated lysine as described for Panel A.