A SENSITIVE RADIOISOTOPIC METHOD FOR THE MEASUREMENT OF NAD(P)H - ITS APPLICATION TO THE ASSAY OF METABOLITES AND ENZYMATIC-ACTIVITIES

被引：70

作者：

SENER, A ^{[1
]}

MALAISSE, WJ ^{[1
]}

机构：

[1] FREE UNIV BRUSSELS,EXPTL MED LAB,B-1050 BRUSSELS,BELGIUM

来源：

ANALYTICAL BIOCHEMISTRY | 1990年 / 186卷 / 02期

关键词：

D O I：

10.1016/0003-2697(90)90073-I

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. © 1990.

引用

页码：236 / 242

页数：7

共 7 条

[1]

Lowry O.H., 1972, FLEXIBLE SYSTEM ENZY

[2] STIMULUS-SECRETION COUPLING OF GLUCOSE-INDUCED INSULIN RELEASE - METABOLIC EFFECTS OF MENADIONE IN ISOLATED ISLETS .31. [J].

MALAISSE, WJ ;

HUTTON, JC ;

KAWAZU, S ;

SENER, A .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1978, 87 (01) :121-130

[3]

Malaisse WJ, 1984, METHODS DIABETES RES, V1, P147

[4] DISPERSED RAT PAROTID ACINAR CELLS .1. MORPHOLOGICAL AND FUNCTIONAL CHARACTERIZATION [J].

MANGOS, JA ;

MCSHERRY, NR ;

BUTCHER, F ;

IRWIN, K ;

BARBER, T .

AMERICAN JOURNAL OF PHYSIOLOGY, 1975, 229 (03) :553-559

[5] PERTURBATION OF PANCREATIC-ISLET FUNCTION IN GLUCOSE-INFUSED RATS [J].

MARYNISSEN, G ;

LECLERCQMEYER, V ;

SENER, A ;

MALAISSE, WJ .

METABOLISM-CLINICAL AND EXPERIMENTAL, 1990, 39 (01) :87-95

[6] DETERMINATION OF INTRACELLULAR PYRIDINE-NUCLEOTIDE LEVELS BY BIOLUMINESCENCE USING ANAEROBIC-BACTERIA AS A MODEL [J].

SCHMID, U ;

SCHIMZ, KL ;

SAHM, H .

ANALYTICAL BIOCHEMISTRY, 1989, 180 (01) :17-23

[7] STIMULUS-SECRETION COUPLING OF GLUCOSE-INDUCED INSULIN RELEASE .26. ARE SECRETORY AND FUEL FUNCTIONS OF GLUCOSE DISSOCIABLE BY IODOACETATE [J].

SENER, A ;

PIPELEERS, DG ;

LEVY, J ;

MALAISSE, WJ .

METABOLISM-CLINICAL AND EXPERIMENTAL, 1978, 27 (10) :1505-1517

← 1 →