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Microassays for glucose 6-phosphate and 6-phosphogluconate based on bioluminescent techniques

✍ Scribed by J. Palmisano; J.H. Schwartz

Publisher
Elsevier Science
Year
1982
Tongue
English
Weight
410 KB
Volume
126
Category
Article
ISSN
0003-2697

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✦ Synopsis

Microassays for glucose 6-phosphate (G6P) and 6phosphogluconate (6PG) capable of measurements in the picomole range are described. In these assays NADP+ reduction is coupled stoichiometrically to the oxidation of G6P or 6PG using glucosed-phosphate dehydrogenase or 6-phosphogluconic dehydrogenase, respectively. The NADPH produced by these redox reactions is measured by bioluminescent technique using NADPH-specific bacterial luciferase. Problems encountered with these assays are detailed. To demonstrate the application of these methods for measurements in biological tissues, measurements of G6P and 6PG levels in mucosal cells from the turtle urinary bladder are presented.

Principles. The G6P and 6PG assays con-FMN, flavin mononucleotide; PCA, perchloric acid; HPLC, high-pressure liquid chromatography.

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