Glucose Uptake Assay Kit ab136955 is a simple and direct colorimetric assay for the measurement of glucose uptake in cell cultures.
- Simple, non-radioactive protocol based on 2-deoxyglucose
- Cited in over 180 publications
- Suitable for both suspension & adherent cells
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Glucose Uptake Assay Kit ab136955 is a simple and direct colorimetric assay for the measurement of glucose uptake in cell cultures.
- Simple, non-radioactive protocol based on 2-deoxyglucose
- Cited in over 180 publications
- Suitable for both suspension & adherent cells
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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 1
Human osteosarcoma cancer cell lines 143B and U2OS 2
Human breast cancer cell lines 3
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) Glucose Uptake Assay Kit (Fluorometric) ab136956
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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.
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.
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.
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Terms & Conditions.
Standard curve and example data. 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.
Glucose uptake in 3T3-L1 adipocytes stimulated with insulin (I). 3T3-L1 adipocytes were differentiated using:
Dexamethasone Dexamethasone, anti-inflammatory glucocorticoid ab120743 (1mM, 1:1000)
IBMX IBMX, Non-specific cAMP and cGMP inhibitor ab120840 (11.5 mg/mL, 1:100)
Insulin Recombinant human Insulin protein (Active) ab123768 (1 mg/mL, 1:1000)
Assay Procedure. Step A: 2-DG oxidation to generate NADPH; Step B: NADPH recycling amplification Reaction.
Zhao et al used Lactate assay kit L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 and Glucose uptake assay kit ab136955 to investigate in cell culture supernatants of mouse primary liver cells and UBR7 over-expressing BEL-7402 cells.
WT and Alb-Cre;UBR7fl/fl mouse primary liver cells were analysed for glucose uptake and lactate secretion levels. Glucose uptake and lactate secretion levels were detected in UBR7 overexpressing BEL-7402 cells. Data are shown as mean +/- SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.
Lactate assay kit (L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330) and Glucose Uptake Assay (ab136955) were purchased from Abcam. The detection of lactate, glucose uptake and ATP levels were carried out according to the method recommended by the kit.
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