Description
Key facts
Detection method:Colorimetric,
Sample types:Suspension cells, Adherent cells,
Assay type:Cell-based (quantitative),
Sensitivity:<= 0.01 nmol/well,
Assay time:3h,
Assay Platform:Microplate reader
Product details:
How the assay works
Glucose Uptake Assay Kit (Colorimetric) is an assay that uses the glucose analog 2-deoxyglucose (2-DG) to detect and quantify glycose uptake in cells. 2-deoxyglucose (2-DG) is used in glucose uptake assay protocol 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 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 transfer supernatant to new tubes
- Add supernatants and standards to wells
- Add reaction mix A and incubate for 1 hr at 37°C
- Add extraction buffer and heat to 90°C 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 37°C
How other researchers are using Glucose Uptake Assay ab136955
Glucose Uptake Assay Kit (Fluorometric) has been used in a variety of sample types including:
Human cervical epithelium HeLa and Human neuroblastoma SH-SY5Y
Human osteosarcoma cancer cell lines 143B and U2OS
Human breast cancer cell lines
References:
1-Son S et al. 2023
2-Mei Z al. 2024
3-Peng Z et al. 2024
Related and recommended products
For fluorometric detection, we recommend Glucose Uptake Assay Kit (Fluorometric)
ab136956
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
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 uptake often referred to as "glucose transport" or "glucose direct" is the process by which cells absorb glucose from the bloodstream. This process requires specific transporter proteins primarily the glucose transporter family known as GLUTs. GLUTs differ in their affinities and tissue distributions; for example GLUT4 found in adipose tissues and muscle is insulin-sensitive. Glucose uptake mainly occurs in tissues with high energy demands like muscles and the brain ensuring these tissues receive adequate energy for function and development.
Biological function summary
The role of glucose uptake in cellular energy metabolism is paramount. It supplies glucose as a primary energy substrate especially in tissues that depend heavily on glycolysis. In cells glucose uptake does not act alone but is part of a complex metabolic carbohydrate mechanism. Beyond energy metabolism this process supports biosynthetic pathways and contributes to the maintenance of cellular function and survival. An example of glucose uptake importance includes energy provision during periods of growth or recovery highlighting its role in diverse biological contexts.
Pathways
The process of glucose uptake integrates into critical biological pathways like glycolysis and the broader glucose metabolism. It starts with the activation of insulin signaling pathways which then stimulate glucose transporter translocation to the cell membrane. Proteins such as insulin receptor substrates (IRS) and PI3K/Akt are intimately linked to these pathways facilitating signal transduction that triggers glucose uptake. These pathways not only regulate glucose levels in the body but also influence processes such as protein synthesis and cell growth.
Glucose uptake malfunctions are closely associated with diabetes and metabolic syndrome. Reduced sensitivity to insulin leads to impaired GLUT4 translocation and therefore decreased glucose uptake in muscle and adipose tissues elevating blood glucose levels— a hallmark of type 2 diabetes. Moreover cancer cells exhibit altered glucose uptake where overexpression of certain GLUTs like GLUT1 allows them to satisfy increased metabolic demands thereby promoting proliferation. Aberrations in these mechanisms highlight the critical connection between glucose uptake and disease pathogenesis.
