An NADH-linked luciferase assay for glycogen: the preparation of glycogen-free, viable human eccrine sweat glands.
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.
Welch
C
C
Department of Clinical Biochemistry, University of Newcastle upon Tyne, United Kingdom.
Bryant
A
A
Kealey
T
T
D000255:Adenosine Triphosphate / Q000032:analysis; D001419:Bacteria / Q000201:enzymology; D004439:Eccrine Glands / Q000032:analysis*; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000032:analysis; D006003:Glycogen / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008156:Luciferases; D008722:Methods; D009243:NAD; D013545:Sweat Glands / Q000032:analysis*
D009243:NAD; D000255:Adenosine Triphosphate; D006003:Glycogen; D005948:Glucose Dehydrogenases; D042843:Glucose 1-Dehydrogenase; D008156:Luciferases