Lane 1: Wild-type HCT116 cell lysate (20µg)
Lane 2: CDKN1A knockout HCT116 cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab109199 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109199 Anti-p21 antibody [EPR3993] was shown to specifically react with p21 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266860 (knockout cell lysate ab256870) was used. Wild-type and p21 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109199 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.