5'-Atp → L-Tyrosine

Participants

Reaction Steps

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.

Provenance & Evidence