ab75974 staining p57 Kip2 in HeLa cells treated with dexamethasone (ab120743), by ICC/IF. Increase in p57 Kip2 expression correlates with increased concentration of dexamethasone as described in literature.The cells were incubated at 37°C for 24h in media containing different concentrations of ab120743 (dexamethasone) 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 ab75974 (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.
1e7 3T3-L1 cells were treated for 15 minutes with 1 µM dexamethasone (ab120743) and 1 µg x mL-1 insulin (ab123768) with the addition of 50 µM forskolin (ab120058), 0.5 mM IBMX (ab120840) or 0.1 mM 8-CPT-cAMP (ab120424), extracted according to the protocol. Lysates were diluted 50-fold, and tested in duplicates (+/-SD).