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1 14c glucose

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1-14C-glucose is a radiochemical product used in research applications. It is a form of the glucose molecule that is labeled with the radioactive carbon-14 isotope at the first carbon position. This product can be used as a tracer compound in various scientific studies and experiments.

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Lab products found in correlation

6 14c glucose, by PerkinElmer (6 mentions) Ls6000 liquid scintillation analyzer, by Beckman Coulter (3 mentions) Ready safe liquid scintillation fluid, by Beckman Coulter (2 mentions) Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions) Whatman filter paper, by Cytiva (1 mentions) Ls6500 scintillation counter, by Beckman Coulter (1 mentions)

Close spelling variants of this product

2-deoxy-D-[1-14C] glucose 2-7deoxy-D-[1-14C]glucose (2-[14C]DG) [1-14C]-2-deoxy-glucose 2-deoxy-D-[1-14C]-glucose (2-DOG) 1-14C glucose and 6-14C glucose D-[1-14C]-glucose (Glu∗) D-(1-14C(U))glucose D-[1-14C] glucose 2-deoxy-D-[1-14C]glucose (2-[14C]DG) 14C-glucose

8 protocols using 1 14c glucose

1

Glucose Oxidation Assay with Antimalarials

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One million cells were plated onto 10 cm dishes in triplicate, grown overnight and treated with vehicle control (water) or with CQ or Q at the indicated doses for 3h at 37C. Cells were washed twice with PBS and the tissue culture media was replaced with media containing 0.8μCi of 1-14C-glucose (Perkin Elmer) or 6-14C-glucose containing vehicle (water) or the indicated antimalarial; cells were incubated for an additional 7h prior to analysis. To capture gaseous 14CO2, Whatman filter paper was taped to the inside of culture dish lid and the plate was then sealed with Parafilm; the experiment was terminated by adding 500μL of perchloric acid to the cells; the plate was resealed and left in the incubator overnight. Filter papers were then carefully removed and placed into vials containing Ready-Safe Liquid Scintillation Fluid (Beckman Coulter), and 14CO2 activity was measured on a Beckman LS6000 Liquid Scintillation Analyzer. The incubation of cells with 6-14C-glucose, used to correct for 14CO2 production from the citric acid cycle, yielded minimal 14CO2 activity.
Salas E., Roy S., Marsh T., Rubin B, & Debnath J. (2015). Oxidative Pentose Phosphate Pathway Inhibition Is A Key Determinant of Antimalarial Induced Cancer Cell Death. Oncogene, 35(22), 2913-2922.
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2

Measuring Glucose Oxidation via the PPP

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Glucose oxidation via the PPP was measured as previously described based on the difference in 14CO2 production from [1-14C] glucose (decarboxylated in the 6-phosphogluconate dehydrogenase-catalyzed reaction and in the Krebs cycle) and [6-14C] glucose (decarboxylated only in the Krebs cycle) (Cisternas et al., 2014 (link)). Slices were washed with ice-cold PBS and collected by trypsinization. The tissue was then resuspended in an O2-saturated Krebs Henseleit buffer, and 500 μl of this suspension (~106 cells) was placed in Erlenmeyer flasks with another 0.5 ml of Krebs Henseleit solution containing 0.5 μCi D-[1-14C] glucose or 2 μCi D-[6-14C] glucose and 5.5-mM D-glucose (final concentration). The Erlenmeyer flasks were equipped with a central well-containing an Eppendorf tube with 500 μl of benzethonium hydroxide. The flasks were flushed with O2 for 20 s, sealed with rubber caps, and incubated for 60 min in a 37°C water bath with shaking. The incubations were stopped by the addition of 0.2 ml of 1.75-M HClO4 into the main well, and shaking was continued for another 20 min to facilitate the trapping of 14CO2 by benzethonium hydroxide. Radioactivity was quantified as previously described by liquid scintillation spectrometry (Bolaños et al., 2008 (link); Herrero-Mendez et al., 2009 (link)). Both [1-14C] glucose and [6-14C] glucose were purchased from PerkinElmer (Waltham, MA, USA).
Cisternas P., Gherardelli C., Salazar P, & Inestrosa N.C. (2021). Disruption of Glucose Metabolism in Aged Octodon degus: A Sporadic Model of Alzheimer's Disease. Frontiers in Integrative Neuroscience, 15, 733007.
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3

Glucose Metabolism Tracing via 14C Labeling

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After washing in PBS, cells were detached with PBS containing 2.5% v/v FBS and 2 mmol/L EDTA, rinsed with PBS, and resuspended in 1 mL Hepes buffer (145 mmol/L NaCl, 5 mmol/L KCl, 1 mmol/L MgSO4, 10 mmol/L Hepes, 10 mmol/L glucose, 1 mmol/L CaCl2, pH 7.4). 50 μL were taken up, sonicated and used to measure the protein content. In each remaining sample, 2 μCi of [6-14C] glucose (55 mCi/mmol, PerkinElmer) or 2 μCi of [1-14C] glucose (58 mCi/mmol, PerkinElmer) were added. The labelled cell suspension was incubated for 1 h in a closed tube to trap the 14CO2 produced from the [14C] glucose. After this incubation time, the metabolic flux was interrupted by adding 0.5 mL 0.8 N HClO4 [17 (link)]. [1-14C] glucose metabolized through PPP or TCA and [6-14C] glucose metabolized through the TCE only produced 14CO2. The amount of glucose producing CO2 through the PPP was obtained by subtracting the amount of [6-14C] glucose (TCA cycle) from [1-14C] glucose (TCA + PPP cycles), as described [17 (link)]. Results were expressed as nmol CO2/h/mg cell proteins.
Roux C., Mucciolo G., Kopecka J., Novelli F., Riganti C, & Cappello P. (2021). IL17A Depletion Affects the Metabolism of Macrophages Treated with Gemcitabine. Antioxidants, 10(3), 422.
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4

Glucose Metabolism Quantification in Cells

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Cells were washed with fresh medium, detached with trypsin/EDTA (0.05/0.02% v/v), washed with PBS, and resuspended in 1 ml Hepes buffer (145 mmol/L NaCl, 5 mmol/L KCl, 1 mmol/L MgSO4, 10 mmol/L Hepes, 10 mmol/L glucose, 1 mmol/L CaCl2, pH 7.4) containing 2 μCi of [6-14C] glucose (55 mCi/mmol, PerkinElmer) or 2 μCi of [1-14C] glucose (58 mCi/mmol, PerkinElmer). A 50 μL aliquot was sonicated and used for the determination of the cell proteins. The remaining suspension of cells was incubated for 1 h in a closed experimental system to trap the 14CO2 produced from the [14C] glucose, and the reaction was stopped by injecting 0.5 mL 0.8 N HClO4, as described previously (42). 14CO2 is released when [1-14C] glucose is metabolized, either by the PPP or by the TCA, whereas it is only developed from [6-14C] glucose only via the TCA. The amount of glucose transformed into CO2 through the PPP was calculated as described [42 (link)] and expressed as nmol CO2/h/mg cell proteins.
Capello M., Ferri-Borgogno S., Riganti C., Chattaragada M.S., Principe M., Roux C., Zhou W., Petricoin E.F., Cappello P, & Novelli F. (2015). Targeting the Warburg effect in cancer cells through ENO1 knockdown rescues oxidative phosphorylation and induces growth arrest. Oncotarget, 7(5), 5598-5612.
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5

Quantifying Cellular Lipogenesis via 14C Glucose

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For measurement of lipogenesis, cells were starved in no-glucose serum-free media for 24 h. Following starvation, labeling with 1-14C glucose (PerkinElmer) was performed overnight. Cells were washed twice with PBS before lysis in 0.5% Triton X-100. The lipid fraction was extracted by the addition of chloroform and methanol (2:1 v/v). Samples were centrifuged, and 14C incorporation was measured from the lipid-containing phase using a scintillation counter. Each condition was normalized to total cellular protein concentrations and assessed using a Bicinchoninic Acid Protein Assay Kit (ThermoFisher Scientific).
Shinde A.B., Nunn E.R., Wilson G.A., Chvasta M.T., Pinette J.A., Myers J.W., Peck S.H., Spinelli J.B, & Zaganjor E. (2023). Inhibition of nucleotide biosynthesis disrupts lipid accumulation and adipogenesis. The Journal of Biological Chemistry, 299(5), 104635.
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6

Measuring Oxidative Pentose Phosphate Pathway

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To measure oxidative PPP, the indicated cell types were plated onto 6 cm dishes, grown for 2d, washed twice with PBS, and the tissue culture media was incubated with media containing 0.8μCi of 1-14C-glucose (Perkin Elmer) for 5h. To capture gaseous 14CO2, Whatman filter paper was taped to the inside of culture dish lid and the plate was then sealed with Parafilm; the experiment was terminated by adding 500μL of perchloric acid to the cells; the plate was resealed and kept at room temperature overnight. Filter papers were then carefully removed and placed into vials containing Scintisafe Plus 50% cocktail scintillation fluid (Fisher Scientific) and 14CO2 activity was measured on a Beckman LS6000 Liquid Scintillation Analyzer. Notably, the incubation of cells with 6-14C-glucose, used to correct for 14CO2 production from the citric acid cycle, yielded minimal 14CO2 activity. To measure relative PPP via NMR, cells were maintained in DMEM containing 2-13C glucose for 18h. The next day, media was collected and 13C spectra acquired on above-mentioned 600 Mz INOVA spectrometer.
Roy S., Leidal A.M., Ye J., Ronen S.M, & Debnath J. (2017). Autophagy-Dependent Shuttling of TBC1D5 Controls Plasma Membrane Translocation of Glut1 and Glucose Uptake. Molecular cell, 67(1), 84-95.e5.
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7

Measuring Pentose Phosphate Pathway Activity

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PPP activity was analyzed as described [27 (link)]. Briefly, cells in 96-well plates were supplemented with 5 μCi/ml of [1-14C]-glucose (specific activity 45-60 mCi/mmol) or [6-14C]-glucose (specific activity 50-62 mCi/mmol, all from PerkinElmer) as indicated in a final volume of 100 μl. The wells were overlaid with Ba(OH)2 impregnated 3MM Whatman paper to capture released 14CO2 in an incubator at 37oC, 5% CO2 for 3 hrs. The paper was removed and placed in an acetone bath, air-dried and incubated at 110 ℃ for 5 minutes. Pieces of the paper corresponding to each well were cut and placed in scintillation counting vials containing scintillation fluid. Their radioactivity was measured in a Beckman LS6500 scintillation counter. The radioactivity of 14CO2 from [1-14C]-glucose was used as a measurement of the carbon flux through the PPP enzyme, 6-phosphogluconate dehydrogenase. PPP-dependent CO2 production was calculated as the difference between 14CO2 derived from [1-14C]-glucose and 14CO2 derived from [6-14C]-glucose (TCA cycle-dependent CO2 production from glucose).
Wang J., Duan Z., Nugent Z., Zou J.X., Borowsky A.D., Zhang Y., Tepper C.G., Li J.J., Fienh O., Xu J., Kung H.J., Murphy L, & Chen H.W. (2016). Reprogramming metabolism by histone methyltransferase NSD2 drives endocrine resistance via coordinated activation of pentose phosphate pathway enzymes. Cancer letters, 378(2), 69-79.
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8

Glucose Oxidation Assay with Antimalarials

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One million cells were plated onto 10 cm dishes in triplicate, grown overnight and treated with vehicle control (water) or with CQ or Q at the indicated doses for 3h at 37C. Cells were washed twice with PBS and the tissue culture media was replaced with media containing 0.8μCi of 1-14C-glucose (Perkin Elmer) or 6-14C-glucose containing vehicle (water) or the indicated antimalarial; cells were incubated for an additional 7h prior to analysis. To capture gaseous 14CO2, Whatman filter paper was taped to the inside of culture dish lid and the plate was then sealed with Parafilm; the experiment was terminated by adding 500μL of perchloric acid to the cells; the plate was resealed and left in the incubator overnight. Filter papers were then carefully removed and placed into vials containing Ready-Safe Liquid Scintillation Fluid (Beckman Coulter), and 14CO2 activity was measured on a Beckman LS6000 Liquid Scintillation Analyzer. The incubation of cells with 6-14C-glucose, used to correct for 14CO2 production from the citric acid cycle, yielded minimal 14CO2 activity.
Salas E., Roy S., Marsh T., Rubin B, & Debnath J. (2015). Oxidative Pentose Phosphate Pathway Inhibition Is A Key Determinant of Antimalarial Induced Cancer Cell Death. Oncogene, 35(22), 2913-2922.
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