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Glucose Uptake Assay Kit (Fluorometric) (CAT#: CB-P163-K) Datasheet

Product Type
Kit
Description
The glucose uptake assay kit (Fluorometric) is a highly sensitive and easy-to-use non-radioactive kit that can detect glucose uptake as low as 50 pmol/well in various cell types.
Since 2-deoxyglucose (2-DG) is similar in structure to glucose, it is used in the glucose uptake measurement protocol. 2-DG is absorbed by glucose transporter and metabolized to 2-DG-6-phosphate (2-DG6P). 2-DG6P cannot be further metabolized, so it will accumulate in the cell. The accumulated 2-DG6P is directly proportional to the 2-DG (or glucose) uptake by the cells. In this experiment, the accumulated 2-DG6P enzyme is oxidized and coupled to a probe, which fluoresces in the presence of NADPH.
Applications
Functional Studies
Size
1 kit
Detection Method
Colorimetric/Fluorometric
Sensitivity
= 0.05 nmol/well
Sample
Adherent cells, Suspension cells
Species Reactivity
Mammals
Protocol Length
2h 00m
Components
2-Deoxyglucose 1 x 1ml
2-DG Uptake Assay Buffer 1 x 10ml
2-DG6P Standard (Lyophilized) 1 vial
Enzyme Mix (lyophilized) 1 vial
Extraction Buffer 1 x 17ml
Neutralizing Buffer 1 x 1ml
PicoProbe 1 x 0.2ml
Requires
Microplate reader
Handling Advice
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 optionally 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
Storage
Please refer to protocols.
Shelf Life
12 months
Research Use
For research use only. Not for human or drug use

Figure 1 Glucose Uptake measured in 3T3-L1 Adipocytes; I = Insulin.


Figure 2 Standard curve: mean of duplicates (+/- SD) with background reads subtracted


Figure 3 Assay Procedure

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All products and services are for Research Use Only. Do Not use in humans.

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