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Non-Radioactive Glucose Uptake Assays
Glucose transport systems are responsible for transporting glucose across cell membranes. Measuring the uptake of 2 deoxyglucose (2-DG), a glucose analog, is widely accepted as a reliable method to estimate the amount of glucose uptake and to investigate the regulation of glucose metabolism and insulin resistance. The 2-DG uptake is commonly determined using the non-metabolized 2-DG labeled with tritium or C14. However, the use of a radiolabeled probe is costly and requires a tedious special handling procedure.
Fig. 1
Glucose transportation
Glucose transportation into cells via glucose transporters, Glu 1, Glu 2, Glu 3 and Glu 4.
AAT Bioquest's Screen Quest™ Colorimetric Glucose Uptake Assay Kit provides a sensitive and non-radioactive glucose uptake assay. 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 cells and is proportional to glucose uptake. The accumulated 2-DG6P is enzymatically coupled to generate NADPH, which is specifically monitored by a chromogenic NADPH sensor. The signal can be read using an absorption microplate reader by reading the OD ratio of 570 nm to 610 nm.
Fig. 2
2-DG uptake
2-DG uptake in differentialed 3T3-L1 adipocytes and 3T3-L1 fibroblasts. Assays were performed with Screen Quest™ Fluorimetric Glucose Uptake Assay Kit in a black wall/clear bottom cell culture Poly-D-Lysine plate using a Gemini (Molecular Devices) microplate reader. A: Negative Control, no insulin and no 2-DG treatment; B: 2-DG uptake in the absence of insulin; C: 2-DG uptake in the presence of 1 µM insulin; D: 2- DG uptake in the presence of 1 µM insulin and 200 µM phloretin; E: 2-DG uptake in the presence of 1 µM insulin and 5 mM D-glucose.
Based on the same principle, AAT Bioquest's Screen Quest™ Fluorimetric Glucose Uptake Assay Kit provides an even more sensitive and non-radioactive glucose uptake assay in tissues or cultured cells. The accumulated 2-DG6P is enzymatically coupled to generate NADPH, which is specifically monitored by a fluorogenic NADPH sensor. The signal can be read using a fluorescence microplate reader at Ex/Em = 540 nm/590 nm.

Document: 03.0027.130501r1
Last updated Thu Oct 02 2025
Non-Radioactive Glucose Uptake Assays