Lane 1: Wild-type HEK293T cell lysate (20µg)
Lane 2: SQSTM1 knockout HEK293T cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab56416 observed at 62 kDa. Red - loading control ab181602 observed at 37 kDa.
ab56416 Anti-SQSTM1 / p62 antibody - Autophagosome Marker was shown to specifically react with SQSTM1 in wild-type HEK293T cells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type and SQSTM1 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. ab56416 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4^°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Lane 1: Wild-type HEK293T cell lysate (20µg)
Lane 2: SQSTM1 knockout HEK293T cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab109012 observed at 64 kDa. Red - loading control ab8245 observed at 37 kDa.
ab109012 Recombinant Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker was shown to specifically react with SQSTM1 in wild-type HEK293T cells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type and SQSTM1 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. ab109012 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 10000 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.