An enzymatic fluorimetric assay to quantitate 2-deoxyglucose and 2-deoxyglucose-6-phosphate for in vitro and in vivo use

Previously, we developed a microplate assay to quantitate 2-deoxyglucose (2DG) and 2-deoxyglucose-6-phosphate in samples for in vitro and in vivo use. In this assay system, four different reaction mixtures were used, and the difference in the reactivity of the two types of glucose-6-phosphate dehydrogenase (G6PDH) variants was used. Because G6PDH from tolura yeast was no longer available, we modified our assay system for the use of G6PDH from Leuconostoc. Using this improved assay system, concentrations of glucose, 2DG, glucose-6-phosphate, and 2-deoxyglucose-6-phosphate were easily measured. This assay may be useful for measuring uptake of 2DG without the use of radioisotopes. (C) 2010 Elsevier Inc. All rights reserved.