Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme

✍ Scribed by P.E. Stanley; S.G. Williams

• Publisher: Elsevier Science
• Year: 1969
• Tongue: English
• Weight: 708 KB
• Volume: 29
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(69)90323-6

✦ Synopsis

The mechanism of action of the ATP-dependent luciferin-luciferase enzyme which gives rise to bioluminescence is now well understood (l-3). This enzyme, extracted from the firefly (Photinus pyrulis) , has been used extensively for the assay of ATP (4-9) since it is the most sensitive and specific method known. Some compounds and enzymes which are conjugate in their action with ATP have also been assayed using this system (10-12). Of the methods described, two basic approaches are evident:

1. In the first one the intensity of the initial light flash is measured with a photometer.

2. The second method uses a quantum counter such as a liquid scintillation spectrometer to measure the number of photons produced over a definite time interval after adding the enzyme to the vial in the laboratory (7,8). No modification of commercially available instruments is needed and with current models it is possible to measure as little as, 1CF mole ATP.

This paper examines factors affecting the use of a liquid scintillation spectrometer for the assay of ATP. A simple, rapid method has been developed for determining ATP over a wide range (10-O to lo-l2 mole) using a single setting on the spectrometer. In addition it is shown that perchloric acid extracts containing ATP may be assayed directly without neutralization.