The first 4 residues GPHP are from Turbo3C cleavage site.
Phosphorylated at S602.
Component and Formulation:
PAK7KD: 1 mg/ml in Storage Buffer of 25 mM Tris-HCl pH 8.0, 150 mM NaCl, 10% glycerol, 5 mM DTT.
Activity:
Specific activity:4,199 pmoles/min/µg
Analysis of enzymatic activity was performed according to the Zlyte assay protocol (Invitrogen):
Different concentrations of PAK7KD were incubated in a buffer containing 50 mM HEPES (pH7.5), 10 mM MgCl2, 1 mM EGTA, 200 mM ATP, 0.01% Brij-35, and 2 µM substrate (SER/THR 14, Invitrogen) at room temperature for 1 hour.
Developer solution was added to the reaction and the reaction was stopped after 1 hour of incubation at room temperature.
Fluorescence was detected using lexc=460±40 nm and lem=528±20 nm filters.