製品名Glucose Uptake Assay Kit (Colorimetric)
Glucose uptake キット 製品一覧
サンプルの種類Adherent cells, Suspension cells
検出感度<= 0.01 nmol/well
種交差性交差種: Other species, Mammals
Glucose Uptake Assay Kit (Colorimetric) (ab136955) is a highly sensitive and easy to use non-radioactive assay kit which can detect glucose uptake as low as 10 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 2-DG6P is oxidized to generate NADPH, the level of which can be determined by an enzymatic recycling amplification reaction.
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 heat at 85ºC for 40 min
- cool on ice for 5 min
- add neutralizing buffer, spin and transfer supernatant to new tubes
- add supernatants and standards to wells
- add reaction mix A and incubate for 1 hr at 37ºC
- add extraction buffer and heat to 90ºC for 40 min
- cool on ice for 5 min and add neutralizing buffer
- add reaction mix B
- analyze every 2-3 mins on microplate reader in kinetic mode at 37ºC
Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
保存方法Store at -20°C. Please refer to protocols.
内容 ラベル 100 tests 2-Deoxyglucose Purple 1 x 1ml 2-DG6P Standard (Lyophilized) Yellow 1 vial Assay Buffer WM 1 x 25ml Enzyme Mix (Lyophilized) Orange 1 vial Extraction Buffer NM 1 x 17ml Glutathione Reductase (Lyophilized) Green 2 vials Neutralizing Buffer Clear 1 x 2.5ml Recycling Mix(Lyophilized) Blue 1 vial Substrate Red 2 vials
2-DG6P Standard curve (a) and 2-DG uptake in 3T3-L1 cells (b), Human adipocytes (c) and HeLa cells (d) respectively. I=Insulin; P=Phloretin.
Step A: 2-DG oxidation to generate NADPH; Step B: NADPH recycling amplification Reaction.
This product has been referenced in:
- Li H et al. Fibroblast growth factor 21 increases insulin sensitivity through specific expansion of subcutaneous fat. Nat Commun 9:272 (2018). Read more (PubMed: 29348470) »
- Wu D et al. HIF-1a is required for disturbed flow-induced metabolic reprogramming in human and porcine vascular endothelium. Elife 6:N/A (2017). Read more (PubMed: 28556776) »