Description
Key facts
Detection method:Fluorescent,
Sample types:Adherent cells, Suspension cells,
Assay type:Quantitative,
Sensitivity:= 0.05 nmol/well,
Assay time:2h,
Assay Platform:Microplate reader
Product details:
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 37°C
- wash cells with PBS to remove exogenous 2-DG
- lyse cells with extraction buffer and repeated pipetting
- freeze/thaw lysates and 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
This product is manufactured by BioVision, an Abcam company and was previously called K666 Glucose Uptake Fluorometric Assay Kit. K666-100 is the same size as the 100 test size of ab136956.
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 < 10 pmol/well.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
REACH authorisation
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glucose often referred to as blood sugar is a simple sugar and an essential carbohydrate in biology. It has a molecular mass of 180.16 g/mol and is highly soluble in water. Glucose circulates in the bloodstream and is absorbed by tissues mainly liver muscle and adipose tissue. It serves as a critical energy source and cells use glucose uptake processes to transport glucose across their membranes. Various diagnostic tools and kits such as glucose assay kits and glucose test kits help measure glucose levels in biological samples.
Biological function summary
Glucose serves as the primary energy substrate for cells providing energy through glycolysis and oxidative phosphorylation. It is not part of any protein complexes but it interacts with numerous enzymes and proteins to regulate metabolic processes. Glucose operates in maintaining homeostasis and the brain relies on it almost exclusively for energy. Glucose assay reagents and glucose detection kits are utilized to quantify glucose concentrations in research studies examining these functions.
Pathways
Glucose is a central component in glycolysis and the tricarboxylic acid (TCA) cycle. In glycolysis glucose is broken down into pyruvate generating ATP and NADH in the process. This pathway involves key regulatory proteins such as hexokinase and phosphofructokinase. In the TCA cycle glucose metabolites further produce ATP and CO2 involving enzymes like citrate synthase. Glucose uptake assays provide insights into how these pathways operate under various physiological conditions.
Glucose regulation and metabolism are tightly linked to diabetes mellitus and metabolic syndrome. Diabetes mellitus is characterized by impaired glucose uptake and insulin regulation often involving insulin receptor pathways. Persistent high glucose levels lead to complications such as neuropathy and retinopathy. AMP-activated protein kinase (AMPK) plays an important role in metabolic syndrome by affecting glucose uptake and energy balance. Understanding glucose's role in these diseases is central to devising therapeutic strategies and interventions.
