KYSE510 cells were harvested at 24 post-transfection and were counted. 105 cells were incubated with 2 mL glucose-free RPMI-1640 medium in a well of a 6-well cell culture plate for 16 under aforementioned conditions, followed by incubation in high-glucose RPMI-1640 medium under the same cell culture conditions for further 24 h. Finally, intracellular glucose levels were measured using a fluorescence-based glucose assay kit (BioVision, Milpitas, CA, USA).
Fluorescence based glucose assay kit
Manufactured by Abcam
Sourced in United States
The Fluorescence-based glucose assay kit is a laboratory equipment product designed to measure glucose levels in biological samples. It utilizes a fluorescence-based detection method to quantify glucose concentration accurately and reliably.
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15 protocols using fluorescence based glucose assay kit
1
Intracellular Glucose Quantification
2
Intracellular Glucose Quantification via 2-NBDG Assay
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RCC4 cells were transduced with a lentivirus encoding control shRNA or Set7 shRNAs. Intracellular glucose level was determined with a fluorescence-based glucose assay kit (BioVision).
2-NBDG glucose uptake assays were also performed for MEFs and RCC4 cells. In brief, the cells were grown under specified conditions with or without 100 μM 2-NBDG (Invitrogen) in the media for 1 h. Glucose uptake was quantified by flow cytometry (FACS), as previously described (64 (link),66 (link)).
2-NBDG glucose uptake assays were also performed for MEFs and RCC4 cells. In brief, the cells were grown under specified conditions with or without 100 μM 2-NBDG (Invitrogen) in the media for 1 h. Glucose uptake was quantified by flow cytometry (FACS), as previously described (64 (link),66 (link)).
3
Intracellular Glucose Quantification
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Caski cells expressing control shRNA or E6 shRNA were cultured in glucose-free DMEM for 16 h, and then incubated with high-glucose DMEM under normoxic or hypoxic conditions for an additional 24 h. The culture medium was removed, and the intracellular glucose levels were measured using a fluorescence-based glucose assay kit (BioVision, Milpitas, CA, USA) according to manufacturer’s instructions.
4
Intracellular Glucose Measurement
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U2OS and MG-63 cells were harvested at 24hrs post-transfection. Cells (105) were first cultivated in glucose-free Eagle’s minimum essential medium under condition of 37°C, 95% humidity and 5% CO2 for 16 hrs, followed by cell culture in high-glucose Eagle’s minimum essential medium for additional 24 hrs under the same conditions. After that, a fluorescence-based glucose assay kit (BioVision) was used to measure the levels of intracellular glucose.
5
Glucose Metabolism Analysis in PC Cells
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PC cells were cultured in glucose-free DMEM for 16 h, and then incubated with high-glucose DMEM under normoxic conditions for an additional 24 h. Culture medium was then removed, and intracellular glucose levels were measured using a fluorescence-based glucose assay kit (BioVision, Milpitas, California, USA) according to the manufacturer's instructions. Lactate levels were measured using a lactate oxidase-based colorimetric assay read at 540 nm according to the manufacturer's instructions (Beyotime, Wuxi, China) and normalized to cell number. Pyruvate kinase activity was measured using the Pyruvate Kinase Activity Assay Kit (Jiancheng, China).
6
Glucose Metabolism in CRC Cells
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Lovo cells together with SW480 cells were cultured in a glucose-free DMEM medium for 16 h. The medium was changed by high-glucose DMEM medium and the CRC cells were cultivated for another day. Lactate production and glucose uptake were determined by utilizing a lactate oxidase-based colorimetric assay and a fluorescence-based glucose assay kit (BioVision, Milpitas, CA, USA), respectively.
7
Glucose Metabolism in Colorectal Cancer
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SW480 and LOVO cells were cultivated in glucose-free DMEM medium for 16 h. And then the medium was replaced with high-glucose DMEM medium, and the CRC cells were cultured for a further 24 h. The glucose uptake and lactate production were detected using Fluorescence-based glucose assay kit (BioVision, Milpitas, California, USA) and lactate oxidase-based colorimetric assay.
8
Glucose Metabolism in Transfected HCC
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Transfected HCC cells were cultivated in glucose-free DMEM for 16 h, and the high-glucose DMEM was added to replace the glucose-free DMEM for 24 h. The glucose level of HCC cells was detected with a fluorescence-based glucose assay kit (BioVision, Milpitas, California, USA). Lactate level in the culture medium was examined with a fluorescence-based lactate assay kit (BioVision).
9
Glucose and Lactate Metabolism in PDAC
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Under normoxic conditions, PDAC cells were cultured in DMEM (glucose-free) for 16 h and were then incubated with DMEM (high-glucose) for 24 h. After removing the culture medium, the intracellular glucose levels were then measured by a fluorescence-based glucose assay kit (BioVision, Exton, USA) based on manufacturer’s instructions. Using a lactate oxidase-based colorimetric assay (Beyotime, Wuxi, China), lactate levels were measured according to the manufacturer’s instructions. The cells were plated in a 24-well plate at the density of 2 × 105 cells/ml, and aliquots of media from each well were assessed 24 h later for the amount of lactate present.
10
Quantifying Lactate and Glucose Metabolism
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PANC‐1 and MIA‐PaCa‐2 cells were plated in a 24‐well plate at the density of 2 × 105 cells/mL. Aliquots of media from each well were assessed 24 hours later for the amount of lactate present, using a lactate oxidase‐based colorimetric assay (Beyotime) according to the manufacturer's instructions. For glucose uptake assay, PANC‐1 and MIA‐PaCa‐2 cells were cultured under normoxic conditions in DMEM (glucose‐free) for 16 hours and were then incubated with DMEM (high‐glucose) for 24 hours. Then, culture medium was removed and the intracellular glucose levels were measured by a fluorescence‐based glucose assay kit (BioVision, Exton, PA, USA) according to the manufacturer's instructions.
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