The free glucose of zebrafish larvae was determined using the Glucose Colorimetric/Fluorometric Assay Kit (Sigma-aldrich, St. Louis, MO, USA). Ten larvae were homogenized in 100 μL of glucose assay buffer. The homogenate was cleared by centrifugation and 10 μL of supernatant (equivalent of one larva) was measured [49 (link)]. The reaction was incubated for 30 min at 37 °C according to the manufacturer’s instructions. The optical density was measured at 570 nm in a Microplate Reader (PerkinElmer, Waltham, MA, USA)
Glucose colorimetric fluorometric assay kit
Manufactured by Merck Group
Sourced in United States
The Glucose Colorimetric/Fluorometric Assay Kit is a laboratory equipment used to quantify the concentration of glucose in a sample. It utilizes a colorimetric or fluorometric method to detect and measure the glucose present in the sample.
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Lab products found in correlation
Lactate assay kit, by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Microplate reader, by PerkinElmer (1 mentions)
Glutamine and glutamate determination kit, by Merck Group (1 mentions)
Dialyzed fbs, by Merck Group (1 mentions)
Atp assay kit, by Merck Group (1 mentions)
Trehalase, by Merck Group (1 mentions)
Amyloglucosidase, by Merck Group (1 mentions)
Accu check guide glucometer, by Roche (1 mentions)
6 protocols using glucose colorimetric fluorometric assay kit
1
Zebrafish Larval Glucose Quantification
2
Glucose and Glutamine Uptake Assay
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Glucose uptake and glutamine uptake were determined by a Glucose Colorimetric/Fluorometric Assay Kit (Sigma-Aldrich) and a Glutamine and Glutamate Determination kit (Sigma-Aldrich), respectively, following the manufacturer’s protocols. Briefly, cells with or without siRNA treatment were cultured in DMEM (Thermo Fisher Scientific) supplemented with 10% dialyzed FBS (Sigma-Aldrich) under matrix-attached or matrix-detached conditions for 6 hours. The medium conditioned with or without cells was collected and assayed for glucose and glutamine concentrations to determine glucose uptake and glutamine uptake. Glucose and glutamine uptake per hour were normalized by the total protein of each sample. Lactate secretion was determined by a Lactate Assay Kit (Sigma-Aldrich) following the manufacturer’s protocol. Briefly, the conditioned medium collected above for glucose uptake and glutamine uptake assays was also assayed for lactate secretion. The lactate secretion per hour was normalized by the total protein of each sample. Glucose uptake rate potential for aerobic oxidation (J) was calculated from glucose uptake rate (E) and lactate secretion rate (F) of the same sample based on the formula
3
Glycolysis Metabolism Analysis
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Glucose consumption, lactate production and ATP level were detected to assess glycolysis metabolism using matched commercial kits, including Glucose Colorimetric/Fluorometric Assay Kit (Sigma-Aldrich), Lactate Assay Kit (Sigma-Aldrich) and ATP Assay Kit (Sigma-Aldrich). All procedures were conducted according to the manufacturer’s instructions.
4
Glycogen and Trehalose Quantification in Yeast
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Yeast cultures were grown and harvested at different times, and approximately 100 OD600 of culture was harvested and washed twice with sterile H2O. The pellets were collected and frozen in liquid nitrogen. The pellets were resuspended in 500 μL of 0.25 M Na2CO3 and incubated for 4 h at 95°C, followed by the addition of 0.15 mL of 1 M acetic acid and 0.65 mL of 0.2 M sodium acetate (pH 5.2) to the sample. Glycogen was digested by 70 U/mg amyloglucosidase (10115, Sigma-Aldrich) overnight at 57 °C. Trehalose was digested by 0.05 U/mL trehalase (T8778, Sigma-Aldrich) overnight at 37 °C under constant shaking. After digestion, the supernatants were collected and the glucose concentration derived from glycogen or trehalose was measured using the Glucose Colorimetric/Fluorometric assay kit (MAK263, Sigma-Aldrich).
5
Interstitial Fluid Glucose Measurement
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Xenografts were dissected within 2 min after the mice were sacrificed by cervical dislocation. They were weighed, briefly washed in PBS, put on a cell sieve with 20-μm pores (pluriStrainer, 20 μm, Pluriselect) on top of a 50-ml tube, and centrifuged at 300g at 4°C for 10 min. Flow-through interstitial fluid was collected (5 to 100 μl per xenograft). Glucose concentration in the interstitial fluid was measured by the Glucose colorimetric/Fluorometric Assay Kit (Merck), according to the manufacturer’s instructions. Five microliters of interstitial fluid was used per measurement. Blood glucose was measured by tail vein puncture using an Accu-Check Guide glucometer (Roche).
6
Glucose Concentration Measurement
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The glucose concentration in the cultures’ supernatant was measured with a glucose colorimetric/fluorometric assay kit (Merck) according to the manufacturer's protocol.
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