A1: The amount of enzyme required varies when different substrates are used. Start with 1-2 µl for 1µg of protein for one hour in a 10-25 µl reaction. If there is still unphosphorylated material, let the reaction go overnight.
Q2: What is the consensus sequence for GSK-3 (P6040)?
A2: It is substrate-dependent. In some target proteins, prior phosphorylation is required, but a general recognition sequence has not yet been elucidated.
