mammalian homologue Ste20-like kinase
5-20mU of MST3 (diluted in 50mM Tris (pH 7.5), 0.1mM EGTA) is assayed against MBP in a final volume of 25.5µl containing 50mM Tris (pH 7.5), 0.1mM EGTA, 0.33mg/ml MBP, 10 mM magnesium acetate and 0.02 mM [33P-g-ATP]( 500-1000 cpm/pmole) and incubated for 30 mins at room temperature. Assays are stopped by addition of 5µl of 0.5M (3%) orthophosphoric acid. Assays are harvested onto P81 Unifilter plates using a wash buffer of 50mM orthophosphoric acid.
MST3_5.pdf (84.57 KB)