ab32557 staining p38 (phospho T180 + Y182) in MALME-3M cells treated with betulinic acid (ab120654), by ICC/IF. Increase in expression of p38 (phospho T180 + Y182) correlates with increased concentration of betulinic acid, as described in literature.The cells were incubated at 37°C for 30 minutes in media containing different concentrations of ab120654 (betulinic acid ) 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 ab32557 (1/200 dilution) 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.