Determination of picogram amounts of ATP using the luciferin-luciferase enzyme system.
G. Lyman, J. Devincenzo
Dec 1, 1967
Citations
94
Citations
Quality indicators
Journal
Analytical biochemistry
Full text
Semantic Scholar
Key Takeaway Key Takeaway: This ultramicro technique allows for the determination of ATP amounts from 10 pg to 10 g with less than 7% variability in quantitative luminescent recordings.
Abstract
Abstract An ultramicro technique for the measurement of ATP based on the firefly luminescent reaction has been presented. The system consists of a photomultiplier tube mounted next to a geometrically designed mirror assembly, which concentrates the luminescence onto the light-sensitive area of the tube. The photomultiplier tube relays the light signal to a photometer, and this in turn to an appropriate chart recorder. An electronically timed injection system has been designed to introduce the luciferin-luciferase extract into the test solution. Incubation of firefly enzyme with an ATPase markedly diminishes residual ATP, which must be eliminated before amounts of ATP below the 1 ng level can be determined. With this procedure it is possible to measure ATP throughout the range of 10 pg to 10 μg with less than ±7% variability in the quantitative luminescent recordings.