Partially purified NDRG2 from rabbit skeletal muscle was maximally phosphorylated with SGK1, PKB alpha, RSK1 or S6K1. Each sample was subjected to SDS/PAGE, transferred to ImmobilonP membranes and immunoblotted with the phospho specific antibodies raised against the sites on NDRG2 at a concentration of 1 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).