Anti-AKT2 Antibody - Western Blot. Anti-AKT2 Antibody in Western blot shows detection of AKT2. A lysate from a stable HEK293 cell line expressing an inducible, myristoylated form of Akt2 (MyrAkt2-ER) was loaded for SDS- PAGE, separated and then transferred to nitrocellulose. The blot was used at 1:1000 and reacted with Anti-AKT2 Antibody for 1h at room temperature. In response to 4OHT (tamoxifen), the Akt2 is recruited to the plasma membrane via its myristoylation sequence, and becomes phosphorylated and activated. The blot (right panel) shows 0 m, 15 m, and 30 m of 4OHT treatment. This treatment has no effect on endogenous Akt2, but causes a band shift upwards in the MyrAkt2. Endogenous Akt2 runs at about 60kD whereas the myristoylated construct runs at around 110kD. Antibody to AKT2 detects both unphosphorylated and phosphorylated AKT2. As before approximately 20 mg/lane of crude HEK293 lysate was loaded for SDS-PAGE, separated and then transferred to nitrocellulose. The right lane contains lysate pretreated-for-15min-with 25 uM LY294002 which affects the phosphorylation of endogenous Akt2 but has no effect on phosphorylation of the myristoylated construct. This antibody clearly detects both the phosphorylated (top arrow) and the non-phosphorylated (bottom arrow) forms of endogenous Akt2.