sensitive and non-radioactive assay for direct and rapid measurement of glucose uptake ability in living cells;
the assay is based on direct incubation of mammalian cells with a fluorescent glucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) followed by cytometric detection of fluorescence producing cells;
cells undergo a short-time incubation in a glucose free media (to normalize the glucose uptake rate across groups) followed by a 20-30 min. incubation with 0,1-0,2 mM 2-NBDG. After washing, the cells are analysed in the green fluorescence channel. It is important to note that 2-NBDG assay is an index only measurement for glucose uptake and should not be compared with glycoloitic rate.
Glucose receptor Glut1
Glut1 is exposed in the cell membrane upon request;
Glut1 is recycled;
the total expression of Glut1 or
intracellular traficking involved in Glut1 targeting and recycling.