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MAK083 Sigma-Aldrich

Glucose Uptake Colorimetric Assay Kit

sufficient for 100 colorimetric tests

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Description

General description

Glucose is the primary source of energy for most cells. Glucose uptake into cells is highly regulated and the first rate limiting step in glucose metabolism. Glucose uptake is facilitated by the GLUT family of transporter proteins, whose expression and activity are regulated by multiple mechanisms. Glucose uptake is upregulated in many cancer cells, which exhibit high rates of aerobic glycolysis. Cells exhibiting insulin resistance show diminished glucose uptake in response to insulin stimulation.

The use of the recycling amplification reaction in the colorimetric assay results in the limit of detection being 10-fold lower (20-100 pmole) compared to the fluorescence assay (200-1,000 pmole, MAK084).

Application

Glucose uptake colorimetric assay kit has been used to measure glucose uptake by a variety of cells.

Suitability

Suitable for detecting glucose uptake in adherent or suspension cells cultured in a 96-well microtiter plate.

Principle

The Glucose Uptake Colorimetric Assay kit provides a simple and direct procedure for measuring glucose uptake in a variety of cells. Glucose uptake is measured using the glucose analog, 2-deoxyglucose (2-DG), which is taken up by cells and phosphorylated by hexokinase to 2-DG6P. 2-DG6P cannot be further metabolized and accumulates in cells, directly proportional to the glucose uptake by cells. In this assay, 2-DG uptake is determined by a coupled enzymatic assay in which the 2-DG6P is oxidized, resulting in the generation of NADPH, which is then determined by a recycling amplification reaction in which the NADPH is utilized by glutathione reductase in a coupled enzymatic reaction that produces glutathione. Glutathione reacts with DTNB to product TNB, which is detected at 412 nm.

Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 
RIDADR 
NONH for all modes of transport
Protocols & Articles
Peer-Reviewed Papers
15

References

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