Notes: The Glutamate-Glo™ Assay was used to measure intracellular glutamate levels. Briefly, the cells were seeded in a 96-well plate, followed by glucose starvation as described above. Cells were then washed with ice-cold PBS and lysed with 0.6 N HCl. The lysing reagent was inactivated using 1 M Tris base. Glutamate dehydrogenase uses the glutamate present in the lysate and NAD+ to produce αKG and NADH. In the presence of NADH, a pro-luciferin reductase substrate is converted by reductase to luciferin, which is then used by Ultra-Glo™ recombinant luciferase to produce light. (4998)

Notes: The authors of this paper describe bioluminescent glucose, lactate, glutamine and glutamate assays suitable for high-throughput analysis of two major metabolic pathways in cancer cells: glycolysis and glutaminolysis. (4769)