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DOI: 10.1055/s-0028-1094058
© Georg Thieme Verlag KG Stuttgart · New York
The Measurement of ATP in Pancreatic Islets by Means of the Luciferin-Luciferase Reaction[*]
Publication History
Publication Date:
07 January 2009 (online)

Abstract
This paper describes a modification of the luciferin-luciferase method for the determination of ATP. The reaction was found to be inhibited by glycyl-glycine-HCl, by arsenate and by albumin; therefore, it was carried out in N-2-hydroxyethylpiperazine-N'-2-ethane sulfonic acid (Hepes), a buffer which has the added advantage of not inhibiting luciferase. Sucrose, ascorbic acid, and mercapto-ethanol were used to stabilize the enzyme activity. A glucose dependent increase in the intracellular ATP concentration of pancreatic islets in vitro was shown. The insulinogenic action of glucose appears to be mediated by ATP.
Key words
Luciferase - Isolated Pancreatic Islets - Insulin Secretion - ATP - Cyclic AMP
1 Aided by Grant No. AM 06034 from the National Institute of Arthritis and Metabolic Diseases.
1 Aided by Grant No. AM 06034 from the National Institute of Arthritis and Metabolic Diseases.
2 Trainee, Diabetes Training Grant No. AM 05474, National Institute of Arthritis and Metabolic Diseases.