Lane 1: Wild-type HeLa cell lysate (20µg)

Lane 2: AKT1 knockout HeLa cell lysate (20µg)
Lane 3: MCF7 cell lysate (20µg)
Lane 4: Daudi cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab108202 observed at 60 kDa. Red - loading control ab181602 observed at 37 kDa.
ab108202 Anti-AKT1 antibody [M1843] was shown to specifically react with AKT1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264852 (knockout cell lysate ab256835) was used. Wild-type and AKT1 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. ab108202 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4^°C at 1 in 1000 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.

Allele-1: 1 bp insertion in exon 2

 

Allele-2: Insertion of the selection cassette in exon 2

 

Allele-3: Insertion of the selection cassette in exon 2