Lane 1: Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate 30 ug
Lane 2: Wild-type A549 IFN-y (ab259377) (100 ng/ml, 32 h) and TNF-alpha (ab259410) (10 ng/ml) for 32 hours, and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate 30 ug
Lane 3: IP10 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate 30 ug
Lane 4: IP10 knockout A549 IFN-y (ab259377) (100 ng/ml, 32 h) and TNF-alpha (ab259410) (10 ng/ml) for 32 hours, and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate 30 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab137018 observed at 11 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab137018 was shown to react with IP10 in wild-type A549 cells in western blot with loss of signal observed in IP10 knockout cell line ab266969 (IP10 knockout cell lysate ab256886). A549 wild-type and IP10 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab137018 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4^°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.