Glucose transport systems are responsible for transporting glucose across cell membranes. Measuring uptake of 2-deoxyglucose (2-DG), a glucose analog, in tissues and cells is widely accepted as a reliable method to estimate the amount of glucose uptake and to investigate the regulation of glucose metabolism and mechanism of insulin resistance. The 2-DG uptake is commonly determined by using non-metabolized 2-DG labeled with tritium or C14. However, routine use of a radiolabelled probe is costly and requires a tedious special handling procedure. AAT Bioquest’s Screen Quest Fluorimetric Glucose Uptake Assay Kit provides a sensitive and non-radioactive assay in tissues or cultured cells. In this assay 2-DG is taken up by glucose transporters, and metabolized to 2-DG-6-phosphate (2-DG6P). The non-metabolizable 2-DG6P accumulates in the cells, and is proportional to glucose uptake by cells. The accumulated 2-DG6P is enzymatically oxidized and generates NADPH, which is specifically monitored by a fluorogenic NADPH sensor. The signal can be read by a fluorescence microplate reader at Ex/Em = 540 nm/590 nm.