反应详情

An Abl reaction mixture was prepared by mixing 1 ml of reaction buffer (50 mM Tris-HCl, 10 mM MgCl2 1 mM EGTA 2 mM dithiothreitol (pH 7.5 at 25° C.)), 10 μl of 10 mM ATP, 2 μl of 6-(9-oxo-9H-acridin-10yl) hexanoate-labelled Abl Peptide substrate (non-phosphorylated) (500 μM in DMSO). 100 μl of this mixture was placed into the wells of a black flat bottomed microtitre plate. The reaction was initiated by the addition of 10 μl of Abl kinase (New England Biolabs, Code P6050L Lot 5), (100,000 units/ml or 100 units/μl) which had been diluted in reaction buffer (50 mM Tris-HCl, 10 mM MgCl2, 1 mM EGTA, 2 mM dithiothreitol, 0.01% Brij 35 (pH 7.5 at 25° C.) to a concentration of 100 units per 10 μl.

1. customAn Abl reaction mixture
2. customwas prepared
3. additionby mixing
4. additionThe reaction was initiated by the addition of 10 μl of Abl kinase (New England Biolabs, Code P6050L Lot 5), (100,000 units/ml or 100 units/μl) which