Glucose Uptake-Glo™ Assay
The Glucose Uptake-Glo™ Assay is a non-radioactive, plate-based, homogeneous bioluminescent method for measuring glucose uptake in mammalian cells based on the detection of 2-deoxyglucose-6-phosphate (2DG6P). When 2-deoxyglucose (2DG) is added to cells, it is transported across the membrane and rapidly phosphorylated in the same manner as glucose. However, enzymes that further modify glucose-6-phosphate (G6P) cannot modify 2DG6P, and thus this membrane-impermeable analyte accumulates in the cell. After a brief period of incubation, an acid detergent solution (Stop Buffer) is added to lyse cells, terminate uptake and destroy any NADPH within the cells. A high-pH buffer solution (Neutralization Buffer) is then added to neutralize the acid. A Detection Reagent containing glucose-6-phosphate dehydrogenase (G6PDH), NADP+, Reductase, Ultra-Glo™ Recombinant Luciferase and proluciferin substrate is added to the sample wells. G6PDH oxidizes 2DG6P to 6-phosphodeoxygluconate and simultaneously reduces NADP+ to NADPH. The Reductase uses NADPH to convert the proluciferin to luciferin, which is then used by Ultra-Glo™ Recombinant Luciferase to produce a luminescent signal that is proportional to the concentration of 2DG6P.
Features - Benefits
- Use a Non-Radioactive Assay: The assay is based on the same principal as the radioactive approach, but no radioactivity is required.
- Follow a Simple and Homogeneous Protocol: After addition of 2DG, there are no wash steps—all steps are additions.
- Achieve Sensitivity with Broad Linearity: The Glucose Uptake-Glo™ Assay can detect 0.5 to 30µM 2DG6P and generates a signal-to-background ratio >3 with as few as 5,000 cells.
- Automate your Workflow: The add-and-read format is compatible with automated and high-throughput workflow; reactions are scalable for use in 96- and 384-well plates.
- Get Reliable and Reproducible Results: The Glucose Uptake-Glo™ Assay yields Z´ factors >0.5.
Assay glucose uptake in mammalian cells, including insulin-sensitive cell types and cancer cells.
Using 50µl of sample + 25µl of Stop Buffer + 25µl of Neutralization Buffer + 100µl of 2DG6P Detection Reagent in a 96-well plate: Cat.# J1341 provides sufficient reagents to perform 50 assays; Cat.# J1342, 100 assays; Cat.# J1343, 500 assays.
Keyword: Glucose update, non-radioactive glucose uptake, glucose metabolism, energy metabolism, oncology, 대사 질환, 당뇨병, 비만
- Customer type - Academia, Pharma/Biotech
- Application - Metabolism, diabetes, oncology, cell differentiation (iPS)