A facile radiometric technique for measuring ATP

✍ Scribed by David A. Cooney; H.N. Jayaram; Elton R. Homan; Charles F. Motley

• Publisher: Elsevier Science
• Year: 1974
• Tongue: English
• Weight: 483 KB
• Volume: 62
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(74)90376-5

✦ Synopsis

A facile radiometric technique for measuring ATP in samples of biologic origin is presented. The n-glucose-6-phosphate produced by the phosphorylation of excess tritiated D-glucose with crystalline hexokinase is stoichiometrically equivalent to the ATP present. Product is selectively precipitated with ethanolic barium acetate, washed with ethanol and the tritium label counted in a scintillation spectrometer. The technique is especially suitable for the measurement of ATP in large numbers of samples (ScrlOO) and offers acceptable sensitivity down to 62 fmoles.

Many techniques are available

for measuring ATP (l-5). In the present communication we describe a method with sensitivity and facility not offered by other published methods. Large numbers of samples can be processed with ease and precision, and without the use of special or modified equipment.

Methods

Eppendorf 1500 ~1 polyethylene centrifuge vessels were purchased from Brinkman Instruments, Inc., 8120 Fenton Street, Silver Spring, MD 20910. Hexokinase from yeast, (obtained as a 10 mg/ml crystalline suspension in 3.2 M ammonium sulfate, pH 6, with a sp act of 140 units/mg protein) and o-glucose-6-phosphate were the products of Boehringer, 219 E.