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A modified direct phosphate assay for studying ATPases

โœ Scribed by W. Terry Jenkins; Millie M. Marshall

Publisher
Elsevier Science
Year
1984
Tongue
English
Weight
446 KB
Volume
141
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

A simple, rapid assay for purified ATPases is presented, based upon the formation of phosphomolybdate and its extraction into butyl acetate. The inclusion of imidazole makes the assay more sensitive and reproducible apparently because of the formation of an imidazole-phosphomolybdate complex. Protein (100 micrograms), Hepes buffer [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] (0.1 M) and nucleotides (1 mM) were all shown to cause interference. The interference by nucleotides could be counteracted by using more molybdate. Butyl acetate was shown to extract virtually all of the phosphomolybdate almost instantaneously upon vortex mixing.

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