title: Radioisotopic assay of femtomole quantities of total adenine nucleotides, ATP plus ADP, and AMP.
abstract: AMP is converted to ATP by incubating overnight with pyruvate kinase, phosphoenolpyruvate and adenylate kinase in the presence of endogenous ATP (ADP) as primer. In a subsequent incubation in the presence of pyruvate kinase, phosphoenolpyruvate, radioactive glucose and hexokinase, ATP and ADP are estimated together by coupling their recycling to the formation of glucose 6-phosphate. The latter is separated by precipitation using 76% (v/v) acetone for radioactivity measurement in the same Eppendorf tube. The sensitivity of these simple procedures matches or exceeds those of luciferase methods of nucleotide determination.
mesh_terms: D000227:Adenine Nucleotides / Q000032:analysis*; D000244:Adenosine Diphosphate / Q000032:analysis; D000249:Adenosine Monophosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002250:Carbon Radioisotopes; D066298:In Vitro Techniques; D008832:Microchemistry / Q000379:methods; D008930:Mitochondria, Liver / Q000032:analysis; D051381:Rats
chemical_list: D000227:Adenine Nucleotides; D002250:Carbon Radioisotopes; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate