Rapid protein kinase assay using phosphocellulose-paper absorption.
نویسندگان
چکیده
Protein kinase (EC 2.7.1.37) catalyzes the phosphorylation of serine and threonine residues of a number of proteins. Histone is widely used as an acceptor substrate in measuring the activity of this enzyme isolated from a variety of sources. We have devised a rapid procedure for resolving phosphohistone from ATP and its metabolites based on the specific absorption of phosphorylated histone onto phosphocellulose paper. Using [y-“P]ATP as the phosphoryl donor. aliquots of the protein kinase assay mixture are applied to phosphocellulosepaper disks that are then immersed in water which elutes [y-3’P]ATP and metabolites. After brief organic solvent extraction and drying. bound radioactivity is measured by liquid scintillation spectrometry.
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عنوان ژورنال:
- Analytical biochemistry
دوره 66 1 شماره
صفحات -
تاریخ انتشار 1975