Determination of cyclic nucleotide phosphodiesterase activity by high-performance liquid chromatography.
A new assay for cyclic nucleotide phosphodiesterase activity by high-performance liquid chromatography with on-line radiochemical detection has been developed. The method is based on the measurement of 3H-labeled nucleoside monophosphates formed from cyclic nucleotides by the action of 3',5'-cyclic-nucleotide phosphodiesterase (PDE). The reaction products are determined from the incubation mixture after removal of the protein by injection of an aliquot into the liquid chromatograph. The detection limit with counting efficiency of 30% is 20 fmol of 3H-labeled product, which makes the method suitable for detection of low PDE activities.
D015056:1-Methyl-3-isobutylxanthine; D000818:Animals; D002147:Calmodulin / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology
D002147:Calmodulin; D007202:Indicators and Reagents; D015056:1-Methyl-3-isobutylxanthine