A sensitive radioisotopic method for the measurement of NAD(P)H: its application to the assay of metabolites and enzymatic activities.
A radioisotopic method for the assay of NADH or NADPH is presented, which is based on the conversion of 2-[U-14C]ketoglutarate to 14C-labeled glutamate in the reaction catalyzed by glutamate dehydrogenase. The efficiency of the method is close to 75%, its precision (coefficient of variation) close to 5%, and its sensitivity close to 0.1 pmol/sample. This simple and rapid method can be applied to the measurement of several metabolites and enzymatic activities. In the present study, its application to the assay of sorbitol, 3-hydroxybutyrate, glutamate dehydrogenase, 3-hydroxybutyrate dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase is documented.
Sener
A
A
Laboratory of Experimental Medicine, Brussels Free University, Belgium.
Malaisse
W J
WJ
D020155:3-Hydroxybutyric Acid; D000818:Animals; D002250:Carbon Radioisotopes / Q000032:analysis; D005069:Evaluation Studies as Topic; D005969:Glutamate Dehydrogenase / Q000032:analysis; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis; D006884:Hydroxybutyrate Dehydrogenase / Q000032:analysis; D006885:Hydroxybutyrates / Q000032:analysis; D009249:NADP / Q000032:analysis*; D051381:Rats; D013012:Sorbitol / Q000032:analysis
D002250:Carbon Radioisotopes; D006885:Hydroxybutyrates; D013012:Sorbitol; D009249:NADP; D006884:Hydroxybutyrate Dehydrogenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005969:Glutamate Dehydrogenase; D020155:3-Hydroxybutyric Acid