Sodium Fluoride → Tromethamine hydrochloride

Participants

Reaction Steps

The competion experiments were carried out in the following way: The standard reaction mixture contained 2.5 mM MgCl2, 10 mM dithio threitol, 1 mg/ml bovine serum albumin, 2.5 mM ATP, 10 mM NaF, 2 μM methyl-3H-thymidine, an appropriate amount of a nucleoside analog and the multisubstrate deoxyribonucleoside kinase in an amount resulting in a linear conversion of the substrate, in a total reaction mixture of 50 μl of 50 mM Tris-HCl, pH 8.0. The reaction mixture was incubated at 37° C. for 30 min, and the reaction was terminated by spotting on to DE-81 discs (Whatman). The discs were instantly immersed and washed in ethanol (70%). The filters were dried and assayed for radioactivity in a tuluene-based scintillant. The concentration resulting in 50% inhibition of phosphorylation of 2 μM deoxythymidine was determined as a mean value of three independent experiments.

Provenance & Evidence