Lane 1: Wild-type HCT116 cell lysate (20µg)
Lane 2: PODXL knockout HCT116 cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab150358 observed at 200 kDa. Red - loading control ab7291 observed at 50 kDa.
ab150358 Anti-PODXL antibody [EPR9518] was shown to specifically react with PODXL in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266887 (knockout cell lysate ab257211) was used. Wild-type and PODXL 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. ab150358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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.