Glucose Uptake Assay Kit (Fluorometric) (ab136956)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 2 hr
- Sample type: Adherent cells, Suspension cells
- Sensitivity: 0.05 nmol/well
Overview
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Product name
Glucose Uptake Assay Kit (Fluorometric)
See all Glucose kits -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells -
Assay type
Quantitative -
Sensitivity
= 0.05 nmol/well -
Assay time
2h 00m -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
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 37ºC
- wash cells with PBS to remove exogenous 2-DG
- lyse cells with extraction buffer and repeated pipetting
- freeze/thaw lysates and optionally heat at 85ºC 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 37ºC -
Notes
If you want a more sensitive assay, we recommend using Glucose Uptake Assay Kit (Colorimetric) (ab136955), which contains an amplification step that allows the kit to detect
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components Identifier 100 tests 2-Deoxyglucose Purple 1 x 1ml 2-DG Uptake Assay Buffer WM 1 x 10ml 2-DG6P Standard (Lyophilized) Yellow 1 vial Enzyme Mix (lyophilized) Green 1 vial Extraction Buffer NM 1 x 17ml Neutralizing Buffer Clear 1 x 1ml PicoProbe Blue 1 x 0.2ml -
Research areas
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Relevance
Glucose (C6H12O6; FW: 180.16) is a ubiquitous energy source in most organisms, from bacteria to humans. The breakdown of carbohydrates produces mono- and disaccharides, most of which is glucose. Through glycolysis and TCA (citric acid cycle), glucose is oxidized to eventually form CO2 and water, generating the universal energy molecule ATP. Glucose is a primary source of energy for the brain and a critical component in the production of proteins and in lipid metabolism and therefore measurement of glucose level is a key diagnostic parameter for many metabolic disorders.
Images
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Functional Studies - Glucose Uptake Assay Kit (Fluorometric) (ab136956)Glucose Uptake measured in 3T3-L1 Adipocytes; I = Insulin.
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Functional Studies - Glucose Uptake Assay Kit (Fluorometric) (ab136956)Standard curve: mean of duplicates (+/- SD) with background reads subtracted -
Example data2-DG6P Standard curve (a) and 2-DG uptake in Human adipocytes (b) , HeLa Cells (c) and 3T3-L1 cells (d) respectively. I=Insulin; P=Phloretin. -
Assay ProcedureAccumulated 2-DG6P is enzymatically oxidized and coupled to the picoprobe, which generates fluorescence in the presence of NADPH.
