Serine / threonine protein kinase 6
Autotactivated with Mg/ATP
Aurora A (5–20 mU diluted in 50 mM Tris pH 7.5, 0.1 mM EGTA, 0.1% β-mercaptoethanol, 1mg/ml BSA) is assayed against LRRLSLGLRRLSLGLRRLSLGLRRLSLG in a final volume of 25.5µl containing 50mM Tris pH 7.5, 0.1 mM EGTA, 0.3mM LRRLSLGLRRLSLGLRRLSLGLRRLSLG, 10 mM magnesium acetate and 0.005mM [33P-γ-ATP] (50-1000 cpm/pmole) and incubated for 30 min at room temperature. Assays are stopped by addition of 5 µl of 0.5 M (3%) orthophosphoric acid and then harvested onto P81 Unifilter plates with a wash buffer of 50 mM orthophosphoric acid.
Aurora_A.pdf (86.13 KB)