Conformation specific anti rabbit igg

Antibodies to PDH kinase-4 (PDK4), PGC-1α (for immunoprecipitation), and histone H4 were purchased from Santa Cruz Biotechnology (Dallas, TX). Antibodies to acetylated lysine, acetylated p53 (Lys³⁷⁹), phospho-AMPK-α (Thr¹⁷²), AMPK-α, conformation-specific anti-rabbit IgG, and GAPDH were from Cell Signaling Technology (Danvers, MA). Total PGC-1α and Sirt1 were measured using antibodies from Millipore (Billerica, MA). Antibodies against p53, COXIV, and ATP5F1 were purchased from Novus Biologicals (Littleton, CO). Standard immunoprecipitation procedures (Cell Signaling Technology) and Western blotting procedures (Invitrogen, Carlsbad, CA) were performed, with GAPDH used as a loading control for whole-tissue samples; histone H4 protein was used as the loading control for nuclear extracts. Quantitation by densitometry used Scion image software (Frederick, MD), with data expressed as the ratio of band intensity of the protein of interest to loading control. When immunoblotting procedures of protein samples from the AdrKO and AdrTG groups were performed in the same run; the data were expressed as a ratio of the WT in the AdrKO group. When the blotting procedures were performed separately, the WTs in relevant individual groups (AdrKO or AdrTG) were used as the control.