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Chemiluminescent Assay of Co-factors by Akio Tsuji; Masako Maeda; Hidetoshi Arakawa is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Chemiluminescent Assay of Co-factors about?

Bioluminescent methods are widely used for the assay of the co-factors, NADH and ATP. Although the bioluminescent method is highly sensitive, the enzymes used are unstable and expensive. Therefore a chemiluminescent method would be valuable in clinical routine assay. We have developed a chemiluminescent method for the assay of NADH using the I-methoxy-5-methylphenazinium methyl sulphate (1-MPMS)/isoluminol(lL)/microperoxidase(m-POD) system. In order t o increase the sensitivity of this method, enzymatic cycling system was coupled to the chemiluminescent assay of NADH. Alcohol dehydrogenase and malate dehydrogenase were used as the cycling enzyme. The standard curve was obtained in the range from 3 x to 5 x 10-12mol/assay. The detection limit of NADH was 30fmol/assay which was comparable to that of the bioluminescent method using bacterial luciferase. Two chemiluminescent methods for the assay of ATP have been developed. Method 1 is the system using hexokinaseIG6PDH and 1-PMS/IL/m-POD, and method 2 is the system based on the enzymatic cycling reaction of ATP using hexokinasdpyruvate kinase. Method 2 is 1000/fold more sensitive than the method 1. The detection limit of ATP was 10fmoV

Who reads Chemiluminescent Assay of Co-factors?

Author

Akio Tsuji; Masako Maeda; Hidetoshi Arakawa

Publisher

Gale Ecco, Print Editions

Published

1989

Language

EN

ISBN

9781170040164

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)