Before and after the cells were transfected with pGFPN1, pGFPN1/MIR181A2HG, pGFPN1/AKT2 or miRNA mimics, glucose in the cell culture medium was measured using Glucose Uptake Colorimetric assay kit (Sigma-Aldrich; MerckKGaA). Glucose uptake was calculated by the difference before and after the cells were treated. After the cells were treated, the glycogen level was detected using a glycogen assay kit (Nanjing Jiancheng Bioengineering Inc.), and the results were normalized to a standard curve and expressed as μg of glycogen, according to the manufacturer's instructions.
Glucose uptake colorimetric assay kit
Manufactured by Merck Group
Sourced in United States, China, Germany
The Glucose Uptake Colorimetric Assay Kit is a laboratory equipment product that measures the glucose uptake in cells. The kit utilizes a colorimetric method to quantify the amount of glucose transported into the cells.
Automatically generated - may contain errors
Lab products found in correlation
Lactate assay kit, by Merck Group (9 mentions)
L lactate assay kit, by Abcam (3 mentions)
Lactic acid assay kit, by Nanjing Jiancheng (2 mentions)
Bicinchoninic acid bca assay kit, by Thermo Fisher Scientific (2 mentions)
Extraction buffer, by Merck Group (2 mentions)
Neutralization buffer, by Merck Group (2 mentions)
Microplate reader, by Molecular Devices (2 mentions)
Lactate assay kit, by Abcam (2 mentions)
Lactate colorimetric assay kit, by Merck Group (2 mentions)
Glycogen assay kit, by Nanjing Jiancheng (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Bca kit, by Vazyme (1 mentions)
Lactic acid assay kit, by Keygen Biotech (1 mentions)
Oxytherm system, by Hansatech (1 mentions)
Phosphate buffer saline (pbs), by Thermo Fisher Scientific (1 mentions)
Lactate assay kit 2, by Merck Group (1 mentions)
Atp detection kit, by Solarbio (1 mentions)
Human insulin, by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Wortmannin, by Merck Group (1 mentions)
41 protocols using glucose uptake colorimetric assay kit
1
Glucose Uptake and Glycogen Measurement
2
Glucose Uptake and Lactate Production
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After the indicated transfection, T98 and U251 cells were cultured in 96-well plates for 48 h. Then cells were washed and collected for analyses of glucose consumption using Glucose Uptake Colorimetric Assay Kit (Sigma) and lactate production using Lactate Assay Kit (Sigma) according to the manufacturer’s instructions. The concentrations of glucose and lactate were analyzed according to the absorbance with a microplate reader (Bio-Rad, Hercules, CA, USA) and normalized to total protein detected by BCA Kit (Vazyme, Nanjing, China).
3
Glucose Uptake Assay Protocol
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Approximately 24 h before the glucose uptake experiments, cells were plated in a 96-well plate at a density of 2000 cells/well. Cells were washed three times with PBS and then glucose-starved by plating in 100 μL of Krebs-Ringer-Phosphate-HEPES (KRPH) buffer composed of 20 mM HEPES, 5 mM KH2PO4, 1 mM MgSO4, 1 mM CaCl2, 136 mM NaCl and 4.7 mM KCl, pH 7.4 containing 2% BSA for 40 min. Glucose uptake was measured using the Glucose Uptake Colorimetric Assay Kit (MAK083, Sigma-Aldrich) according to the manufacturer's instructions. All the experiments were performed in triplicate and repeated three times.
4
Glucose Uptake and Lactate Assay
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A549 and H1299 cells were seeded into 6-well plates and transfected with or without si-KLF5 si-NC, pcDNA-KLF5 or Vector, followed by incubation under a normoxic or hypoxic condition. The culture supernatant of cells was collected after transfection for 48 h and the amount of glucose and lactate in the supernatant was measured using a glucose uptake colorimetric assay kit (Sigma) and a Lactic Acid assay kit (KeyGen, Nanjing, China), respectively.
5
Lactate and Glucose Metabolism Analysis
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Lactate production and glucose consumption were examined using Lactate Assay Kit (Sigma-Aldrich, St. Louis, MO, USA) and Glucose Uptake Colorimetric Assay Kit (Sigma-Aldrich).
6
Glucose and Lactate Measurement in Cell Cultures
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After the treatments as aforementioned, the culture medium of THP-1 and HL60 cells was collected for the measurement of glucose and lactate levels using a glucose uptake colorimetric assay kit (Sigma-Aldrich) and a lactic acid assay kit (Jiancheng Bioengineering Institute, Nanjing, China), respectively.
7
Quantifying Cellular Metabolism in Vitro
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Cells were seeded in 96-well plates at a density of 2000 cells per well. After washing thrice with PBS, glucose uptake was assessed using the Glucose Uptake Colorimetric Assay Kit (Sigma, MAK083, USA). Lactate secretion in the cell culture medium was quantified using the Lactate Assay Kit (Sigma, MAK065, USA). Both assays' results were normalized to the total protein content of each sample. The pH of the medium was measured with a pH meter (Bohlertech Technology, China), and the oxygen consumption rate (OCR) of the cells was determined using the Hansatech Oxytherm system (Hansatech, UK). These experiments were performed in triplicate and replicated three times to ensure consistency.
8
Glucose Uptake Assay in β-like Cells
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Approximately 100 hPSC-derived β-like cells were picked into each well of a 12-well cell culture plate and rinsed three times with PBS. They were then starved in 1 ml DMEM medium with no glucose (GibcoTM) for 4 h. The medium was removed and the β-like cells were rinsed three times with PBS. The cells were then exposed to DMSO or 30, 60, 90 or 120 μM fisetin and incubated in 1 ml DMEM medium containing 1 g/L D-glucose for 4 h before supernatants were collected. The cells were lysed using M-PER protein extraction reagent and measured for protein content. The supernatants were then measured for residual glucose amount using Glucose Uptake Colorimetric Assay Kit (Sigma–Aldrich). The level of glucose uptake was normalized to the protein content in each sample.
9
Glucose Metabolism in Liver Cells
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SMMC-7721 and HepG2 cells were seeded in 96-well plates overnight. After 72 h, cells were washed three times in phosphate buffer saline (Gibco), and then collected for analyses of glucose consumption and lactate production via using the Glucose Uptake Colorimetric Assay Kit or Lactate Assay Kit (Sigma, St. Louis, MO, USA) according to the manufacturer’s instructions, respectively. The concentrations of glucose consumption and lactate production were measured by using a microplate reader and normalized by the total protein concentration determined via the bicinchoninic acid (BCA) assay kit (Thermo Fisher).
10
Glucose Uptake Measurement in Apoptotic Cells
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LR73 cells were incubated with apoptotic Jurkat cells for 2h, washed 3 times with PBS and incubated with 10 mM 2-deoxygloucs (2-DG), a glucose analog, in glucose free media for 30 min. Following incubation, cells were washed 3 times with PBS and lysed with Extraction Buffer (Sigma Cat#: MAK083). Lysate was frozen/thawed in dry ice/ethanol, and then heated at 85°C for 40 min. Lysate was then cooled on ice for 5 min and then neutralized by Neutralization Buffer (Sigma Cat#: MAK083). Samples were spun down at 13,000x g to remove insoluble fraction and then diluted 10-fold by adding Assay Buffer. Using the lysate, glucose uptake was measured using Glucose Uptake Colorimetric Assay Kit (Sigma). 2-DG 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. 2-DG uptake is determined by a coupled enzymatic assay in which the 2-DG6P is oxidized, resulting in the generation of NADPH, which is then determined by a recycling amplification reaction in which the NADPH is utilized by glutathione reductase in a coupled enzymatic reaction that produces glutathione. Glutathione reacts with DTNB to produce TNB, which was detected at 412 nm as per the manufacturer’s recommendations.
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