Lane 1: Wild-type HCT116 cell lysate 20 ug
Lane 2: TRIB3 knockout HCT116 cell lysate 20 ug
Lane 3: Hap1 cell lysate 20 ug
Lane 4: HeLa cell lysate 20 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab75846 observed at 40 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab75846 was shown to react with TRIB3 in wild-type HCT 116 cells in western blot with loss of signal observed in TRIB3 knockout cell line ab273718 (TRIB3 knockout cell lysate ab275249). HCT 116 wild-type and TRIB3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab75846 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4^°C at a 1 in 10000 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^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.