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. 1983 Mar 15;210(3):717-20.
doi: 10.1042/bj2100717.

A simple assay method for determination of the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP

P T Hawkins, R H Michell, C J Kirk

A simple assay method for determination of the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP

P T Hawkins et al. Biochem J. .

Abstract

A simple and rapid assay method is described for determining the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP. Labelled ATP is incubated with cyclic AMP, cyclic AMP-dependent protein kinase and histone H2A under conditions leading to maximum phosphorylation of the histone. The specific radioactivity of the gamma-phosphate group of the 32P-labelled ATP is calculated from the total amount of [32P]Pi incorporated into a standard amount of histone. This assay method uses inexpensive commercially available materials, and it yields an accurate specific radioactivity with as little as 0.25 nmol of 32P-labelled ATP.

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