Extracellular acidification rate (ECAR) was measured using extracellular flux analyzer (XFp) analyzer (Seahorse Bioscience) as described by the manufacturer. Lactate production (Lactate Assay Kit) was measured as described by the manufacturer (BioVision). To measure glucose uptake, cells were incubated with a fluorescent D-glucose derivative, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] -2-deoxy-D-glucose (2-NBDG; APExBIO) for 20 min at 37 °C. The fluorescence intensity of 2-NBDG was measured using by flow cytometry (BD FACSCanto II™), and data were analyzed with FlowJo software (Treestar). ATP production (Enhanced ATP Assay Kit) was measured according to the protocol recommended by manufacturer (Beyotime).
2 nbdg
Manufactured by Apexbio
Sourced in United States
2-NBDG is a fluorescent glucose analog that can be used to measure cellular glucose uptake. It is a non-radioactive, cell-permeable molecule that is transported into cells via glucose transporters and can be detected using fluorescence microscopy or flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Lactate assay kit, by Abcam (3 mentions)
Enhanced atp assay kit, by Beyotime (3 mentions)
2 nbdg, by BD (3 mentions)
M205 fca microscope, by Leica (2 mentions)
2 nbdg, by Beyotime (2 mentions)
Lsrfortessa x 20 flow cytometer, by BD (2 mentions)
Lactic acid assay kit, by Nanjing Jiancheng (2 mentions)
Facscanto 2, by BD (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Glycerol assay kit, by Nanjing Jiancheng (1 mentions)
Atp detection assay kit, by Solarbio (1 mentions)
Chekine lactate assay kit, by Abbkine (1 mentions)
Flow cytometry, by BD (1 mentions)
Spectramax m5 microplate reader, by Molecular Devices (1 mentions)
Guava easycyte 8 flow cytometer, by Merck Group (1 mentions)
2 nbdg, by Merck Group (1 mentions)
Fr180204, by Beyotime (1 mentions)
2 nbdg, by Agilent Technologies (1 mentions)
Fluorescent microscope, by Agilent Technologies (1 mentions)
24 protocols using 2 nbdg
1
Metabolic Profiling of Cell Bioenergetics
2
Glucose Uptake Assay in KGN Cells
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The ability of KGN cells to uptake glucose was evaluated by measuring the fluorescent glucose 2-NBDG (APEXBIO, USA). The KGN cells seeded in 96-well plates were cultured in DMEM/F12 medium. The cells were gently rinsed with DPBS and incubated with 10 μM 2-NBDG at 37 °C for 10 min. The fluorescent intensity of the cells was measured using fluorescence microscopy (Olympus, Japan) with a 450 nm excitation. The level of intracellular fluorescence was quantitatively analyzed using ImageJ software.
3
Glucose Uptake Measurement in HepG2 Cells
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HepG2 cells were seeded in 48-well plates at 1 × 105 cells/well with or without intervention for 48 h at 37°C. After being starved in glucose-free medium for 3 h, the cells were incubated with 40 μM 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy-d -glucose (2-NBDG) (Apexbio; B6035) for 30 min. The 2-NBDG fluorescence intensity was analyzed by flow cytometry at 488 nm (excitation) and 530 nm (emission).
4
Berberine Enhances Glucose Uptake in HepG2 Cells
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HepG2 cells (4 × 104 cells/well) were cultured in a 24-well plate. The 48-h BBR treatment was followed by insulin treatment (100 nM) for 30 min and 0.1 mM 2-NBDG (Apexbio, United States) incubation for 45 min at 37°C. Images were obtained using identical acquisition settings on a fluorescence microscope (Nikon, Japan). Three independent replicates of the experiment were acquired.
5
Glucose Uptake and Metabolite Analysis
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For the glucose uptake assay, 2-NBDG (APExBIO) was used to incubate the cells in the glucose-free medium. Images were then acquired using a fluorescence microscope. Intracellular lactate and ATP levels were determined using a CheKine™ Lactate Assay Kit (Abbkine, Wuhan, China), and ATP detection Assay Kit (Solarbio, Beijing, China), respectively. Glycerol levels of intracellular and tissue were determined using a Glycerol Assay Kit (Nanjing Jiancheng, Nanjing, China). Periodic Acid Schiff (PAS) reagent (Nanjing Jiancheng) was used to detect the Glycogen content according to the manufacturer’s protocol.
6
Quantifying Glucose Uptake Dynamics
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Cells were incubated with ALD-DNA for 0, 6, 12, and 24 h, followed by replacing the medium with a sugar-free medium containing 10% fetal bovine serum and cultured in a carbon dioxide incubator for 30 min. Then, 2-NBDG (APExBIO, Houston, TX, USA) with a final concentration of 100 μM was added and cultured in a carbon dioxide incubator for 15 min. After that, cells were collected and washed twice with PBS and then detected by flow cytometry (BD Biosciences, Dubai, United Arab Emirates) within 10 min.
7
Zebrafish Glucose Uptake Assay
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After the compound treatment, zebrafish (6 dpf) were incubated in a culture medium containing 600 μM 2-NBDG (Apexbio, B6035, Houston, TX, USA) for 3 h. A pool of 5 larvae was homogenized in 100 μL of sample buffer. The homogenate was spun at 10,000 rpm for 10 min. The supernatant (30 μL) was placed into a 96-well plate to detect fluorescence (excitation, blue 475 nm; emission, 500–550 nm) using a SpectraMax M5 Microplate Reader (Molecular Devices). Each sample was measured for three pools [25 (link)].
8
Glucose Uptake Measurement in hESC-ECs
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Glucose uptake was measured using the fluorescence-labeled deoxyglucose analog 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)-2-deoxyglucose (2-NBDG, Apex Bio, USA). After the indicated treatment, hESC-ECs were incubated with 2-NBDG for 1 h at 37 °C, and the fluorescence intensity was measured using the Guava EasyCyte™ 8 flow cytometer (EMD Millipore, Germany).
9
Glucose Uptake Assay in RL95-2 Cells
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To measure glucose uptake, the RL95-2 cells were seeded in 24-well plates (8 × 104 cells) and treated with P4 (0, 10, and 100 nM) for 48 h. The cells were incubated with a fluorescent D-glucose derivative, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (0.2 mM 2-NBDG; APExBIO Technology LLC., Houston, TX, United States) in glucose-free medium for 20 min at 37°C. Images were acquired using a fluorescent microscope, and glucose uptake rate was determined by Image Pro-Plus software and a fluorescein analysis.
10
Zebrafish Glucose Uptake Analysis
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Zebrafish (5 dpf) were incubated in a culture medium containing 600 uM 2-NBDG (Apexbio, B6035, Houston, TX, USA) for 3 h. The larvae were anesthetized for imaging under a M205 FCA microscope (Leica, Wetzlar, Germany), the fluorescence intensity of lens was used as the indicator of glucose uptake according to the reference [32 (link)].
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