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1 2 13c2 glucose

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[1,2-13C2]glucose is a stable isotope-labeled compound that contains two carbon atoms with the 13C isotope. It is used as a research tool in various scientific applications, such as metabolic studies, tracer experiments, and nuclear magnetic resonance (NMR) spectroscopy.

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Lab products found in correlation

U 13c6 glucose, by Cambridge Isotopes (2 mentions) U 13c5 glutamine, by Cambridge Isotopes (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Tributylamine, by Merck Group (1 mentions) U 13c glucose, by Cambridge Isotopes (1 mentions) Optima lc ms grade, by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Phenformin, by Merck Group (1 mentions) Bptes, by Merck Group (1 mentions) U 13c16 palmitate, by Cambridge Isotopes (1 mentions) Poly 2 hydroxyethyl methacrylate, by Merck Group (1 mentions) Palmitic acid, by Merck Group (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) 13c6 glucose, by Cambridge Isotopes (1 mentions) 1 13c1 glucose, by Cambridge Isotopes (1 mentions) 6 13c1 glucose, by Cambridge Isotopes (1 mentions) 3 13c glucose, by Cambridge Isotopes (1 mentions) 5 13c glutamine, by Cambridge Isotopes (1 mentions)

11 protocols using 1 2 13c2 glucose

1

CNS Perfusion Fluid with Isotopic Glucose

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CNS Perfusion Fluid (M Dialysis AB, Stockholm, Sweden) consisted of NaCl (147 mmol/L), KCl (2.7 mmol/L), CaCl2 (1.2 mmol/L), and MgCl2 (0.85 mmol/L) in water. 1,2-13C2 glucose (isotopic enrichment 99%, chemical purity 99%) from Cambridge Isotope Laboratories (Tewksbury, MA, USA) was formulated at 4 or 8 mmol/L in CNS perfusion fluid by the Pharmacy Manufacturing Unit, Ipswich Hospital NHS Trust (Ipswich, UK) who tested the formulations to verify purity, sterility, freedom from endotoxins and absence of pyrogenicity, compliant with current regulations.
Stovell M.G., Howe D.J., Thelin E.P., Jalloh I., Helmy A., Guilfoyle M.R., Grice P., Mason A., Giorgi-Coll S., Gallagher C.N., Murphy M.P., Menon D.K., Carpenter T.A., Hutchinson P.J, & Carpenter K.L. (2023). High-physiological and supra-physiological 1,2-13C2 glucose focal supplementation to the traumatised human brain. Journal of Cerebral Blood Flow & Metabolism, 43(10), 1685-1701.
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2

HPLC-Grade Metabolite Standards

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HPLC-grade water, acetonitrile, and methanol were Optima LC-MS grade obtained from ThermoFisher Scientific (San Jose, CA). The majority of the metabolite standards, as well as tributylamine, acetic acid, and all media components, were obtained through Sigma-Aldrich (St. Louis, MO). U-13C-Glucose (99%) and 1,2-13C2-glucose were obtained from Cambridge Isotope Laboratories (Andover, MA).
Xu Y.F., Lu W, & Rabinowitz J.D. (2015). Avoiding Misannotation of In-Source Fragmentation Products as Cellular Metabolites in Liquid Chromatography–Mass Spectrometry-Based Metabolomics. Analytical chemistry, 87(4), 2273-2281.
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3

Intravenous Glucose Infusion for Traumatic Brain Injury

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At 22 h after their initial surgery (Fig. 1), rats were anesthetized and the right femoral artery and vein were cannulated with PE-50 tubing, as previously described (Bartnik et al., 2007 (link); Bartnik-Olson et al., 2010 (link)). The scalp incision was re-opened, the skull was dried, and the dorsal surface of the skull including the craniotomy site was covered with dental acrylic (to prevent overheating and herniation of cortical tissue through the craniotomy during microwave irradiation). Bupivacaine (0.1–0.14 mg/kg, s.c.) was infiltrated into the femoral and cranial incision sites and animals were restrained on a cardboard plank immediately after removal of isoflurane anesthesia.
At 24 h post-injury awake, lightly restrained animals were given an intravenous infusion of [1,2 13C2] glucose (Cambridge Isotope Laboratories, Andover, MA, USA) in sterile water (Fig. 1). The infusion protocol, per 350 g body weight, began with a bolus injection of 225 μmol followed by 150 μmol given in exponentially decreasing amounts over the next 8 min, until a final constant infusion rate of 16.67 μmol/min was reached and maintained for 1 h (Bartnik-Olson et al., 2010 (link)) (Fig. 1).
Shijo K., Sutton R.L., Ghavim S.S., Harris N.G, & Bartnik-Olson B.L. (2016). Metabolic fate of glucose in rats with traumatic brain injury and pyruvate or glucose treatments: A NMR spectroscopy study. Neurochemistry international, 102, 66-78.
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4

Tracing Glucose Flux through PPP

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A total of 1 × 106 BaF3/ITD or BaF3/ITD-R cells were cultured in a glucose-free RPMI-1640 medium (Gibco, Waltham, MA, USA), supplemented with 11.1 mmol/L [1,2-13C2]-glucose (Cambridge Isotope Laboratories, Tewksbury, MA, USA) and 10% fetal bovine serum (FBS) in 6-well plates for 24 h. Three biological replicates were prepared. Intracellular metabolites were extracted using a methanol/water/chloroform method as previously described [12 (link)]. The extraction, derivatization, and GC–MS analysis procedures were performed as described in above “Pentose phosphate flux analysis with gas chromatography/mass spectrometry” section. Glucose flux through the PPP was calculated as the extracellular glucose uptake flux multiplied by the ratio of (M + 1)/[(M + 1) + (M + 2)], where (M + 1) and (M + 2) are two isotopologues of pyruvate derived from [1,2-13C2]-glucose.
You X., Jiang W., Lu W., Zhang H., Yu T., Tian J., Wen S., Garcia-Manero G., Huang P, & Hu Y. (2019). Metabolic reprogramming and redox adaptation in sorafenib-resistant leukemia cells: detected by untargeted metabolomics and stable isotope tracing analysis. Cancer Communications, 39, 17.
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5

Stable Isotope Metabolic Tracing

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Stable isotopes used in these studies include [1,2-13C2]glucose, [U-13C6]glucose, [U-13C5]glutamine, and [U-13C16]palmitate all acquired from Cambridge Isotope Laboratories. Albumin, phenformin, palmitic acid, and BPTES were acquired from Sigma. Poly(2-hydroxyethyl methacrylate) was acquired from Sigma.
Parker S.J., Svensson R.U., Divakaruni A.S., Lefebvre A.E., Murphy A.N., Shaw R.J, & Metallo C.M. (2016). LKB1 promotes metabolic flexibility in response to energy stress. Metabolic engineering, 43(Pt B), 208-217.
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6

Steady-State 13C Glucose Labeling of Cerebellar Granule Neurons

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Cerebellar granule neurons (7×106 cells per well plated in 2-well Lab-Tek chambers) were incubated for 1 h at 37°C with 0.6 mL experimental buffer containing 9.6 mM [1,2-13C2]glucose (Cambridge Isotope Laboratories, Andover, MA) rather than unlabeled glucose. The cells were further incubated 2 h 15 min at 37°C with 0.8 mL of fresh 1,2 13C glucose buffer (so 3 h 15 min total) to allow the glycolytic, pentose phosphate pathway, and TCA cycle metabolites to reach isotopic steady-state, as previously demonstrated for HepG2 cells (Hofmann et al., 2008 (link); Maier et al., 2008 (link)) and fibroblasts (Munger et al., 2008 (link)). The cells were washed once, then incubated 60 min, 37°C with 0.35 mL of the same buffer. Parallel controls were run in each experiment and included wells without cells, and cells incubated with unlabeled glucose. The labeling pattern of metabolically produced lactate exported during the last hour (from 3 h 15 min to 4 h 15 min of exposure to [13C]glucose) was determined using the supernatants following centrifugation of the samples for 2 min, 21,000 xg, 4°C. The supernatants were stored at −20°C until processed for LC-MS/MS.
Gebril H.M., Avula B., Wang Y.H., Khan I.A, & Jekabsons M.B. (2015). 13C Metabolic flux analysis in neurons utilizing a model that accounts for hexose phosphate recycling within the pentose phosphate pathway. Neurochemistry international, 93, 26-39.
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7

Metabolic Profiling of Murine Cells

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Freshly isolated cells from mice were incubated with RPMI-1640 containing 2% dialyzed FBS (Thermo Scientific, Waltham, MA), 2.0 g/L glucose and cytokines for 16 hours, then replaced in pre-warmed labeling medium containing 2.0 g/L or 0.2 g/L 1,2-13C2-glucose (Cambridge Isotope Laboratories, Andover, MA) with cytokines and incubated for 5 or 60 min at 37°C. Metabolites were extracted with an organic solvent (ice cold 90% 9:1 MeOH: CHCl3) and separated on a 1mm ×150mm HILIC column in a 35 min cycle. All analytes and internal standards were measured by ESI− ionization on an Agilent 6520 QTOF mass spectrometer equipped with a 1260 LC unit (Agilent, Santa Clara, CA).
Saito Y., Chapple R.H., Lin A., Kitano A, & Nakada D. (2015). AMPK protects leukemia-initiating cells in myeloid leukemias from metabolic stress in the bone marrow. Cell stem cell, 17(5), 585-596.
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8

CNS Perfusion Fluid with Isotopic Glucose

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CNS Perfusion Fluid (M Dialysis AB, Stockholm, Sweden) consisted of NaCl (147 mmol/L), KCl (2.7 mmol/L), CaCl2 (1.2 mmol/L), and MgCl2 (0.85 mmol/L) in water. 1,2-13C2 glucose (isotopic enrichment 99%, chemical purity 99%) from Cambridge Isotope Laboratories (Tewksbury, MA, USA) was formulated at 4 or 8 mmol/L in CNS perfusion fluid by the Pharmacy Manufacturing Unit, Ipswich Hospital NHS Trust (Ipswich, UK) who tested the formulations to verify purity, sterility, freedom from endotoxins and absence of pyrogenicity, compliant with current regulations.
Stovell M.G., Howe D.J., Thelin E.P., Jalloh I., Helmy A., Guilfoyle M.R., Grice P., Mason A., Giorgi-Coll S., Gallagher C.N., Murphy M.P., Menon D.K., Carpenter T.A., Hutchinson P.J, & Carpenter K.L. (2023). High-physiological and supra-physiological 1,2-13C2 glucose focal supplementation to the traumatised human brain. Journal of Cerebral Blood Flow & Metabolism, 43(10), 1685-1701.
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9

Isotopic Labeling of TCA Cycle Intermediates

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For the analysis of TCA-cycle intermediates, CSCs were cultured and labeled in the standard serum-free DMEM/F12 media containing 17.5 mM 13C6-glucose (Cambridge Isotope Laboratories). For time course experiments, the secondary-generation mammospheres were cultured in serum-free DMEM/F12 medium for 2 days. The BCSCs were washed three times in medium without glucose, and then the cells were cultured for another 8 h in medium containing 17.5 mM 13C6-glucose. At the time of collection, the cells were washed three times with ice-cold PBS and extracted with 80% methanol on ice. Extracts were dried by centrifugation and stored at −80 °C until further processing. The intermediates were normalized according to internal standard solution containing [U-13C] fumarate, succinate-d4, and citrate-d4 at 1 mM each. For the analysis of lactate through glycolysis, BCSCs were cultured in the medium containing 1,2-13C2-glucose (Cambridge Isotope Laboratories). At the time of collection, the culture medium was centrifuged, transferred to a new Eppendorf tube, and stored at −80 °C until further processing.
Yang D., Peng M., Hou Y., Qin Y., Wan X., Zhu P., Liu S., Yang L., Zeng H., Jin T., Qiu Y., Li Q, & Liu M. (2020). Oxidized ATM promotes breast cancer stem cell enrichment through energy metabolism reprogram-mediated acetyl-CoA accumulation. Cell Death & Disease, 11(7), 508.
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10

Isotope Tracers for Metabolic Analysis

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Tracers ([1-13C1]glucose [98 to 99 atom% 13C], [6-13C1]glucose [99 atom% 13C], [1,2-13C2]glucose [99 atom% 13C], [U-13C6]glucose [99 atom% 13C], [4-2H1]glucose [98 atom% 2H], [5-2H1]glucose [98 atom% 2H], and [1-13C1]ethanol [99 atom% 13C]) were purchased from Cambridge Isotope Laboratories (Andover, MD). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).
Jacobson T.B., Korosh T.K., Stevenson D.M., Foster C., Maranas C., Olson D.G., Lynd L.R, & Amador-Noguez D. (2020). In Vivo Thermodynamic Analysis of Glycolysis in Clostridium thermocellum and Thermoanaerobacterium saccharolyticum Using 13C and 2H Tracers. mSystems, 5(2), e00736-19.
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