The glucose absorption was analyzed with a FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA) by using Glucose Uptake Cell-Based Assay Kit (No.600470, Cayman Chemical). Lactate production was determined using Glycolysis Cell-Based Assay Kit (No.600450, Cayman Chemical) according to the manufacture’s protocol.
Glucose uptake cell based assay kit
Manufactured by Cayman Chemical
Sourced in United States
The Glucose Uptake Cell-Based Assay Kit is a laboratory tool designed to measure glucose uptake in cells. It provides a quantitative method for analyzing cellular glucose absorption and metabolism.
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Lab products found in correlation
Glycolysis cell based assay kit, by Cayman Chemical (5 mentions)
Lactate colorimetric fluorometric assay kit, by Abcam (3 mentions)
Pyruvate assay kit, by Abcam (3 mentions)
Lactic acid assay kit, by Nanjing Jiancheng (3 mentions)
Spectramax m5, by Molecular Devices (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Filtermax f3 f5 multi mode microplate reader, by Molecular Devices (2 mentions)
L lactate assay kit, by Cayman Chemical (2 mentions)
Varioskan flash, by Thermo Fisher Scientific (2 mentions)
Lactate assay kit, by Abcam (2 mentions)
Atp assay kit, by Abcam (2 mentions)
Seahorse xf96 glycolysis analyzer, by Agilent Technologies (2 mentions)
Lactate assay kit wst, by Dojindo Laboratories (2 mentions)
Pdh activity colorimetric assay kit, by Abcam (2 mentions)
Cytoflex, by Beckman Coulter (2 mentions)
Cytoflex flow cytometer, by BD (2 mentions)
Glut1, by Cell Signaling Technology (2 mentions)
Hif 1α, by Cell Signaling Technology (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
82 protocols using glucose uptake cell based assay kit
1
Glucose Uptake and Lactate Production
2
Quantifying Cellular Glucose Consumption
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Glucose consumption was determined in culture media using a glucose assay kit (Sigma-Aldrich, St. Louis, MO, USA). Briefly, cell culture supernatants were harvested and diluted with deionized water, after which 50 μL of the diluted sample was mixed with an equal volume of assay buffer including o-dianisidine in a 96-well plate. The mixture was then incubated at 37 °C for 30 min, at which time the reaction was stopped by adding 50 μL of H2SO4 and the absorbance (O.D.) at 540 nm was measured in a microtiter reader (UVM340, Asys Hitech Gmbh, Eugendorf, Austria). The glucose consumption in each sample was calculated using a calculation formula from a standard curve.
Next, the cellular levels of glucose were measured in C2C12 myotubes using a glucose uptake cell-based assay kit (Cayman Chemical Co., Ann Arbor, MI, USA). Briefly, C2C12 myotubes were treated with or without yam extract and allantoin at different concentrations in glucose-free medium containing 100 µg/mL of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d -glucose (NBDG) for 4 h. After harvesting the cells, cell-based assay buffer (200 µL) was added to each well. The amount of 2-NBDG taken up by the myotubes was then measured with fluorescent filters that detected fluorescein (excitation/emission = 485/650) using a Glomax multi detection system (Promega Biosystems, Sunnyvale, CA, USA).
Next, the cellular levels of glucose were measured in C2C12 myotubes using a glucose uptake cell-based assay kit (Cayman Chemical Co., Ann Arbor, MI, USA). Briefly, C2C12 myotubes were treated with or without yam extract and allantoin at different concentrations in glucose-free medium containing 100 µg/mL of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-
3
Glucose Uptake Assay in Cells
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Glucose uptake assay was performed by using a glucose uptake cell-based assay kit (Cayman, Cat: #600470) according to the manufacturer’s instructions. After cells were cultured with 0.25 mM PA in glucose-free culture medium for 24 h, cells were treated with or without 200 nmol/l insulin for 30 min. Glucose uptake was performed by incubation with a glucose-free culture medium containing 2-NBDG (200 μg/ml) for 30 min and terminated by treatment with cell-based assay buffer before detecting fluorescein (excitation/emission = 485/535 nm).
4
Glucose Uptake Cell-Based Assay
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We assessed cellular glucose uptake using a Cayman Chemical Glucose Uptake Cell-Based Assay Kit following the manufacturer’s protocol (Ann Arbor, MI, USA). Fluorescently labeled deoxyglucose analog 2-NBDG was the labeled glucose utilized for gauging cellular glucose uptake, and fluorescence was analyzed with a microplate reader. In brief, on day 7 of differentiation, cells were washed with PBS, trypsinized, and then transferred to a 96-well black, transparent bottom culture plate. After 24 h, the medium was discarded, and cells were washed with PBS, followed by incubating cells with a mixture of KRPS and 2-NBDG in a 100:1 ratio, and cells were incubated at 37 °C for 2 h. The plate was then centrifuged for 5 min at 400× g at room temperature, and the supernatant was discarded. We then washed cells with PBS and then incubated the cells with Cell-based Assay Buffer before measuring fluorescence under excitation/emission of 485/535 nm.
5
Glucose Uptake Assay in Differentiated C2C12 Cells
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Glucose was measured using glucose uptake cell-based assay kit (Cayman Chemical Co., Ann Arbor, MI, USA). Briefly, C2C12 cells were seeded on a glass plate at 5 × 105 cells/well and then differentiated with DMEM containing 2% horse serum for 5 days. Differentiated cells were then incubated with ARA extract (0.2, 0.5, or 1 mg/mL), metformin (2.5 mM), insulin (100 nM), or apigenin (50 μM) in 1 mL of glucose-free medium containing 150 μg/mL of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d-glucose (NBDG) for 4 h, when supernatants were removed and 200 μL of cell-based assay buffer was added per well. The amount of 2-NBDG taken up by cells was determined by fluorescence microscopy (Leica Biosystems, Wetzlar, Germany).
6
Glucose Uptake in Astrocyte Cultures
7
Quantitative Fluorometric Glucose Uptake Assay
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A glucose uptake cell-based assay kit (Cayman Chemical, Ann Arbor, MI, USA) was used to analyze cellular glucose uptake. Briefly, cells (5 × 104/well) were seeded in 96-well plates and incubated overnight in 100 μl of glucose-free medium, reacted with 200 μg/ml 2-NBDG (2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-d -glucose) for 1 h in glucose-free medium and then centrifuged for 5 min at 400×g at room temperature to remove the supernatant. Two hundred microliters of the assay buffer was added to each well, the plate was centrifuged for 5 min at 400×g at room temperature, the supernatant was aspirated, and 100 μl of assay buffer was added to each well. 2-NBDG fluorescence was quantified using the infinite M200 PRO (TECAN, Männedorf, Switzerland) with excitation/emission wavelengths of 485/525 nm.
8
Fluorescent Glucose Uptake Assay
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Cells (104 per well) were seeded in 96-well plates overnight, and the medium was aspirated and replaced with a fluorescent-tagged glucose derivative (2-NBDG) diluted in glucose-free medium. After an hour, glucose uptake was determined using a Glucose Uptake Cell-based Assay kit (Cayman Chemical, USA) following the manufacturer's instructions. The amount of 2-NBDG taken up by cells was detected by measuring the fluorescein intensity at excitation and emission wavelengths of 485 and 535 nm, respectively. Then, relative glucose uptake was calculated and normalized based on the cell number.
9
Metabolic Profiling of Cultured Cells
10
Ginger Enhances Glucose Uptake in Adipocytes
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To evaluate the insulin-like or insulin-sensitizing effects of ginger on glucose uptake, the fluorescently labeled deoxy-glucose analog, 2-NBDG, was used for the direct detection of glucose taken up in cells using a Cayman Chemical Glucose Uptake Cell-Based Assay Kit (Ann Arbor, MI) as described previously (72 (link)). In brief, 3T3-L1 cells were cultured in 96-well plates at a density of 1 × 104 cells per well, as described above. Cells were then treated with ginger extracts at 0, 25, and 50 μg/mL concentrations for an additional day. The cell culture medium was then changed to glucose-free DMEM medium with or without ginger extracts and further incubated for 8 h. Subsequently, the serum and glucose replete adipocytes were co-cultured with ginger extracts and 2-NBDG at a final concentration of 200 μg/mL for another 16 h. At the end of the treatment, cells were washed twice, and the fluorescence of retained 2-NBDG was measured with a 96-well plate reader (Spectra Max M5, Molecular Devices) at 485 nm (excitation wavelength) and 535 nm (emission wavelength). All experiments were repeated 5 times. The glucose uptake was determined by comparison of 2-NBDG in cells treated with or without ginger extracts.
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