Detection of thymidine kinase activity using an assay based on the precipitation of nucleoside monophosphates with lanthanum chloride.
A new assay method for the measurement of thymidine kinase (TK) is described. Cytosols were prepared from TK- and TK+ cells and evaluated for TK activity using an assay which is based on the phosphorylation of [125I]-iododeoxyuridine, [125I]-iododeoxycytidine, or [3H]thymidine and the precipitation of the monophosphates of these nucleosides by lanthanum chloride. The specificity, reproducibility, sensitivity, and convenience of this assay are demonstrated.
Wolcott
R M
RM
Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130.
Colacino
J M
JM
D001972:Bromodeoxycytidine / Q000031:analogs & derivatives; D002478:Cells, Cultured; D011232:Chemical Precipitation; D003600:Cytosol / Q000201:enzymology; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000378:metabolism; D007065:Idoxuridine / Q000378:metabolism; D007811:Lanthanum / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism; D013937:Thymidine Kinase / Q000032:analysis*
D009711:Nucleotides; C028521:lanthanum chloride; D003841:Deoxycytidine; D001972:Bromodeoxycytidine; C011288:ibacitabine; D007811:Lanthanum; D013937:Thymidine Kinase; D007065:Idoxuridine; D013936:Thymidine