ab39400 staining AMPK alpha 1 (phospho S496) in HepG2 cells treated with AICAR (ab120358), by ICC/IF. Increase in AMPK alpha 1 (phospho S496) expression correlates with increased concentration of AICAR, as described in literature.The cells were incubated at 37°C for 1h in media containing different concentrations of ab120358 (AICAR) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab39400 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.