Glucose Uptake Assay Kit (Fluorometric) ab136956 is a highly sensitive and easy to use non-radioactive kit which can detect glucose uptake as low as 50 pmol/well in a variety of cell types.
2-deoxyglucose (2-DG) is used in glucose uptake assay protocols because of its structural similarity to glucose. 2-DG is taken up by glucose transporters and metabolized to 2-DG-6-phosphate (2-DG6P). 2-DG6P cannot be further metabolized, and thus accumulates within cells. The accumulated 2-DG6P is directly proportional to 2-DG (or glucose) uptake by cells. In this assay, the accumulated 2-DG6P is enzymatically oxidized and coupled to a probe, which generates fluorescence in the presence of NADPH.
Glucose uptake assay protocol summary:
- prepare cells with suitable glucose starvation / uptake stimulation depending on experimental set-up
- add 2-DG to cells and incubate for 20 mins at 37°C
- wash cells with PBS to remove exogenous 2-DG
- lyse cells with extraction buffer and repeated pipetting
- freeze/thaw lysates and heat at 85°C for 40 min
- cool on ice for 5 min
- add neutralizing buffer, spin and retain supernatant
- add supernatants and standards to wells
- add reaction mix and incubate for 40 min at 37°C
This product is manufactured by BioVision, an Abcam company and was previously called K666 Glucose Uptake Fluorometric Assay Kit. K666-100 is the same size as the 100 test size of ab136956.
If you want a more sensitive assay, we recommend using Glucose Uptake Assay Kit (Colorimetric) (Glucose Uptake Assay Kit (Colorimetric)ab136955), which contains an amplification step that allows the kit to detect < 10 pmol/well.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
