We have developed a luminescent glucose uptake assay that is based on the same principle as the aforementioned assays: The accumulation of 2DG6P and detection through the enzymatic action of G6PDH (3). The luminescent assay has a number of advantages:
- Similar sensitivity to the radioactive assay but without any wash steps or handling and disposal of radioactive material.
- Larger signal window than the absorbance and fluorescence assays, making it easier to detect small changes and requiring less assay optimization.
- Amenable to high-throughput screening due to fewer false hits and steady glow-type signals.
The luminescent assay can detect glucose uptake from as few as 5,000 cells with a greater than threefold signal-to-background ratio, and the signal remains linear up to at least 50,000 cells. The luminescent assay is the best combination of simplicity and sensitivity available for a non-radioactive multiwell plate method.