Sener A, Malaisse W J
Laboratory of Experimental Medicine, Brussels Free University, Belgium.
Anal Biochem. 1990 May 1;186(2):236-42. doi: 10.1016/0003-2697(90)90073-i.
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.
本文介绍了一种用于测定NADH或NADPH的放射性同位素方法，该方法基于在谷氨酸脱氢酶催化的反应中，2-[U-14C]酮戊二酸转化为14C标记的谷氨酸。该方法的效率接近75%，精密度（变异系数）接近5%，灵敏度接近0.1 pmol/样品。这种简单快速的方法可用于多种代谢物和酶活性的测量。在本研究中，记录了其在山梨醇、3-羟基丁酸、谷氨酸脱氢酶、3-羟基丁酸脱氢酶和甘油醛-3-磷酸脱氢酶测定中的应用。
