40 μg of the following extracts: rat brain (1), rat testis (2), rat heart (3), rat skeletal muscle (4), human skeletal muscle and HeLa cell were subjected to SDS-Page analysis before being transferred to nitrocellulose membrane and immunoblotted with anti- TORC2 at 0.5 μg/ml. Binding of the primary antibody was detected using rabbit peroxidase conjugated anti- sheep IgG antibody (1 in 10, 000 dilution, Pierce) followed by enhanced chemiluminescence (ECL, Amersham).