ab1793 staining TNFα in RAW 264.7 cells treated with (R)-(-)-rolipram (ab120031), by ICC/IF. Decrease in TNFα expression correlates with increased concentration of (R)-(-)-rolipram, as described in literature.The cells were incubated at 37°C for 24h in media containing different concentrations of ab120031 ((R)-(-)-rolipram) in DMSO, fixed with 100% methanol for 5 minutes at -20°C 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 ab1793 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.