D 5 3h n glucose

Manufactured by PerkinElmer

D-[5-3H(N)]-glucose is a radiolabeled glucose analog used in research applications. It contains a tritium (3H) label at the fifth carbon position of the glucose molecule. This product can be used as a tracer to study glucose metabolism and related processes in biological systems.

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Lab products found in correlation

Ceritinib, by Selleck Chemicals (2 mentions) Ly294002, by Cell Signaling Technology (2 mentions) Rapamycin, by Selleck Chemicals (2 mentions) H2228, by American Type Culture Collection (2 mentions) Mk 2206, by Selleck Chemicals (2 mentions) Cell tak, by BD (1 mentions) D 6 14c glucose, by PerkinElmer (1 mentions) Mitostress kit, by Agilent Technologies (1 mentions) 1 14c palmitic acid, by PerkinElmer (1 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions) Luciferase assay reagent, by Promega (1 mentions) Insulin, by Thermo Fisher Scientific (1 mentions) Epidermal growth factor (egf), by Merck Group (1 mentions) Igf 1, by Thermo Fisher Scientific (1 mentions) Endo free purification kit, by Qiagen (1 mentions) Ag1478, by Merck Group (1 mentions) Fatty acid free bovine serum albumin, by Roche (1 mentions) Anti hk2 antibody, by Cell Signaling Technology (1 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions) 2 deoxy d glucose 2 dg, by Merck Group (1 mentions)

Close spelling variants of this product

[5-3H]-D-glucose [5-3H]glucose D-glucose

6 protocols using d 5 3h n glucose

Investigating Metabolic Regulation by SIRT4

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Bacterial endotoxin [lipopolysaccharide endotoxin (LPS)] was purchased from Sigma (Escherichia coli 0111:B4); d-[5-³H(N)]-glucose, d-[6-¹⁴C]-glucose, 1-[¹⁴C]-palmitic acid and hyamine were obtained from Perkin-Elmer; Mito stress kit was from Seahorse Bioscience; Cell-Tak was from BD company; colorimetric assay kits for pyruvate dehydrogenase (PDH), lactate dehydrogenase (LDH) activities and for pyruvate and lactate levels were from Biovision; gene-specific TaqMan primer/probe sets were from Applied Biosystems; SIRT4 gene-specific siRNA and control siRNA were from Santa Cruz Biotechnology; SIRT4 Flag (Plasmid #13815) (17 (link)), control pcDNA3.1+ without SIRT4 sequence, lentiCRISPRv2 vector, lentivirus helper plasmids psPAX2 and pND2.G were from Addgene; gene-specific antibodies were purchased from Gentex company.

Tao J., Zhang J., Ling Y., McCall C.E, & Liu T.F. (2018). Mitochondrial Sirtuin 4 Resolves Immune Tolerance in Monocytes by Rebalancing Glycolysis and Glucose Oxidation Homeostasis. Frontiers in Immunology, 9, 419.

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Glycolysis Quantification via Tritiated Water

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Glycolysis was measured by the conversion of d-[5-³H(N)]-glucose to tritiated water as described elsewhere (10 (link)). In brief, after starvation with glucose-free RPMI (Gibco) medium, Cells were incubated in triplicates with 1 μCi of d-[5-³H(N)]-glucose (Perkin-Elmer) at 37°C for 1 h in scintillation tubes containing 1 ml of H₂O. The reaction was stopped by adding HCl (1N final) and scintillation tube was sealed. [³H]₂O generated by enolase activity from d-[5-³H(N)]-glucose was vaporized for overnight at 50°C oven and cooled down for another overnight at 4°C. After removal of the central wells, [³H]₂O was counted using a β-counter for detection of glycolytic rate. One μCi of d-[5-³H(N)]-glucose alone in triplicates was set for background control. One μCi of [³H]₂O (Perkin-Elmer) alone in triplicates was set for detecting the efficiency of this water vapor exchange.

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Cell Line Authentication and Culturing

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The cell lines H2228, A549, and A431 were originally obtained from ATCC (Manassas, VA). H3122 was obtained from NCI (Bethesda, MD). All the cell lines were authenticated in July of 2015 by STR DNA fingerprinting at the Characterized Cell Line Core Facility of MD Anderson Cancer Center (Houston, TX). H2228, H3122 and A549 were cultured in RPMI-1640 with 10% fetal bovine serum (FBS). A431 was cultured in DMEM with 10% FBS. D-[5-³H(N)]-glucose was obtained from Perkin Elmer (Boston, MA). Ceritinib, MK2206, and rapamycin were purchased from Selleckchem (Houston, TX). LY294002 was from Cell Signaling (Danvers, MA).

Ma Y., Yu C., Mohamed E.M., Shao H., Wang L., Sundaresan G., Zweit J., Idowu M, & Fang X. (2016). A causal link from ALK to hexokinase II overexpression and hyperactive glycolysis in EML4-ALK-positive lung cancer. Oncogene, 35(47), 6132-6142.

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Cell Line Authentication and Culturing

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Glucose Uptake Regulation by LPA

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LPA (1-oleoly, 18:1) was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL). Prior to use, LPA was dissolved in PBS containing 0.5% fatty acid-free bovine serum albumin (BSA) from Roche (Indianapolis, IN). d-[5-³H(N)]-glucose was purchased from Perkin Elmer (Boston, MA). EGF, 2-deoxy-d-glucose (2-DG), and AG 1478 were obtained from Sigma Aldrich (St. Louis, MO). IGF-1 and insulin were from Invitrogen (Gaithersburg, MD). Plasmid DNAs were purified using the endo-free purification kit from Qiagen (Valencia, CA). Dharmafect 1 was obtained from Dharmacon, Inc. (Lafayette, CO) and TransIT-TKO was obtained from Mirus Bio (Madison, WI). Luciferase assay reagents were obtained from Promega (Madison, WI). Anti-HK2 antibody was obtained from Cell Signaling (Danvers, MA). The TaqMan Universal PCR Master Mix and qPCR probes for HK2 and GAPDH were obtained from Applied Biosystems (Carlsbad, CA).

Mukherjee A., Ma Y., Yuan F., Gong Y., Fang Z., Mohamed E.M., Berrios E., Shao H, & Fang X. (2015). Lysophosphatidic Acid Up-Regulates Hexokinase II and Glycolysis to Promote Proliferation of Ovarian Cancer Cells. Neoplasia (New York, N.Y.), 17(9), 723-734.

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Glucose Flux Measurement in Cells

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This assay was performed as previously published [44] (link). Glucose flux was determined in 2×10⁶ viable cells by washing in PBS followed by incubation in glucose-free Kreb’s buffer for 30 min prior to addition of 10 µCi of D-[5-³H](N)-glucose (PerkinElmer, Wellesley, MA) and non–radio-labeled glucose to bring total glucose concentration to 10 mM prior to culture for 1 hour. Reactions were stopped by addition of an equal volume of 0.2 N HCl. [³H]H₂0 was separated from [³H]glucose by evaporated equilibrium in a sealed environment. Levels of [³H]H₂0 produced were measured on a scintillation counter, and glycolytic flux was calculated as described [44] (link). Average fold change and standard error of the mean were graphed; Student’s two-tailed T-Test was performed to calculate significance at p≤0.05, p≤0.01, or p≤0.001.

Arreola A., Cowey C.L., Coloff J.L., Rathmell J.C, & Rathmell W.K. (2014). HIF1α and HIF2α Exert Distinct Nutrient Preferences in Renal Cells. PLoS ONE, 9(5), e98705.

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