一种用于测定32P标记的ATP的γ-磷酸基团比放射性的简单测定方法。
A simple assay method for determination of the specific radioactivity of the gamma-phosphate group of 32P-labelled ATP.
作者信息
Hawkins P T, Michell R H, Kirk C J
出版信息
Biochem J. 1983 Mar 15;210(3):717-20. doi: 10.1042/bj2100717.
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.
摘要
描述了一种简单快速的测定方法，用于测定32P标记的ATP的γ-磷酸基团的比放射性。将标记的ATP与环磷酸腺苷、环磷酸腺苷依赖性蛋白激酶和组蛋白H2A在导致组蛋白最大磷酸化的条件下孵育。根据掺入标准量组蛋白中的[32P]Pi的总量计算32P标记的ATP的γ-磷酸基团的比放射性。该测定方法使用廉价的市售材料，并且用低至0.25 nmol的32P标记的ATP就能产生准确的比放射性。
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