An NADH-linked luciferase assay for glycogen: the preparation of glycogen-free, viable human eccrine sweat glands

Anal Biochem. 1989 Feb 1;176(2):228-33. doi: 10.1016/0003-2697(89)90300-x.

Abstract

A glycogen assay based on bacterial NADH luciferase is described. It is free of tissue interference. The detection limit is 0.12 nmol glycogen, and the coefficient of variation is 5.5%. A method of depleting human eccrine sweat glands while retaining their viability is described. This depends on their incubation in 10(-5) M acetylcholine and 1 mM pyruvate. This method may be applicable to other tissues. The evidence for the viability of glycogen-depleted human eccrine sweat glands is reported and includes tissue contents of ATP and the rates of oxidation of glucose, pyruvate, beta-hydroxybutyrate, and palmitate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analysis
  • Bacteria / enzymology
  • Eccrine Glands / analysis*
  • Glucose 1-Dehydrogenase
  • Glucose Dehydrogenases / analysis
  • Glycogen / analysis*
  • Glycogen / metabolism
  • Humans
  • Hydrolysis
  • Luciferases*
  • Methods
  • NAD*
  • Sweat Glands / analysis*

Substances

  • NAD
  • Adenosine Triphosphate
  • Glycogen
  • Glucose Dehydrogenases
  • Glucose 1-Dehydrogenase
  • Luciferases