Induction of phosphorylation of STAT 1 at Ser727 in human malignant melanoma cells (short-term culture derived from a patient) in response to interferons. Subconfluent cells were serum-starved before exposure to activation dosages of IFN-gamma (10 ng/ml) and IFN-alpha (1000 IU/ml and 5000 IU/ml). Western blotting analysis of cell extracts shows detection of phosphorylated STAT-1 (Ser727)

Overlay histogram showing HeLa cells stained with ab3987 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3987, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.