How does a glucose uptake assay work?
The glucose uptake assay protocol is based on a fluorescent glucose analog, which just like glucose can be taken up by cells through glucose transporters. However, this glucose analog cannot be fully utilized in glycolysis because of its modification and thus accumulates inside the cells.
How do you measure glucose uptake?
Glucose uptake is measured using the glucose analog, 2-deoxyglucose (2-DG), which is taken up by cells and phosphorylated by hexokinase to 2-DG6P. 2-DG6P cannot be further metabolized and accumulates in cells, directly proportional to the glucose uptake by cells.
What causes glucose uptake?
Insulin increases glucose uptake mainly by enriching the concentration of Glut4 proteins at the plasma membrane, rather than by increasing the intrinsic activity of the transporter (2,3).
What happens during glucose uptake?
Glucose uptake in muscle is a function of different regulatory steps such as delivery of glucose from the blood to the interstitial space, transmembrane transport from the interstitial space to the inside of the muscle cell and intracellular metabolism of the glucose.
What is insulin glucose uptake?
Insulin-Dependent Glucose Uptake Mediated by Subcellular Redistribution of the Glucose Transporter. Insulin is an anabolic hormone that acts on various target tissues, including the liver, skeletal muscle, and fat tissue, regulating the blood glucose level [1,2,3,4].
Which organ depends on insulin for glucose uptake?
Insulin stimulates the liver to store glucose in the form of glycogen. A large fraction of glucose absorbed from the small intestine is immediately taken up by hepatocytes, which convert it into the storage polymer glycogen. Insulin has several effects in liver which stimulate glycogen synthesis.
What is muscle glucose uptake?
Skeletal muscle extracts glucose from the blood to maintain demand for carbohydrates as an energy source during exercise. Such uptake involves complex molecular signalling processes that are distinct from those activated by insulin.
Which cells need insulin for glucose uptake?
Insulin increases glucose uptake into fat and muscle cells through the regulated trafficking of vesicles that contain glucose transporter type 4 (GLUT4).
Is the enzymatic assay useful for measurement of 2DG uptake in insulin-responsive tissues?
Thus, the enzymatic assay should prove to be useful for measurement of 2DG uptake in insulin-responsive tissues in vivo as well as in cultured cells. Copyright © 2011 Elsevier Inc. All rights reserved.
What is a glucose uptake assay kit?
Find out more. 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.
Why does 2-DG inhibit DNA repair?
It has been proposed that exposure to agents that enhance oxidative stress may sensitize cells to the cytotoxic effects of IR. Additionally, 2-DG has been shown to inhibit DNA repair in highly glycolytic cells, such as respiratory-deficient yeast mutants [81].
Does 2-DG interfere with D-mannose?
Thus, 2-DG can interfere in the metabolism of both d-glucose and d-mannose, including inhibiting glycolysis, disrupting mannose-related metabolic pathways, and competing with d-mannose in protein N-glycosylation [47,68,69,70,71].