Lane 1: Wild-type HEK-293T cell lysate 20 ug
Lane 2: AK2 knockout HEK-293T cell lysate 20 ug
Lane 3: HL-60 cell lysate 20 ug
Lane 4: HepG2 cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab166901 observed at 26 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab166901 was shown to react with AK2 in wild-type HEK-293T cells in western blot with loss of signal observed in AK2 knockout cell line ab266539 (AK2 knockout cell lysate ab257825). HEK-293T wild-type and AK2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab166901 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4^°C at a 1 in 1000 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.

Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:AK2 knockout HEK293T cell lysate (20 ug)
Lane 3:HL-60 cell lysate (20 ug)
Lane 4:HepG2 cell lysate (20 ug)

ab157206 was shown to specifically react with AK2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266539 (knockout cell lysate ab257825) was used. Wild-type and AK2 knockout samples were subjected to SDS-PAGE. ab157206 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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.

Allele-1: 8 bp deletion in exon 1

Allele-2: 1 bp insertion in exon 1