2 deoxy d 1 3h glucose

Manufactured by PerkinElmer

Sourced in United States

2-deoxy-d-[1-3H]glucose is a radioactive glucose analog that is commonly used as a tracer in various biological and medical research applications. It functions by allowing researchers to track and measure glucose uptake and metabolism in living cells, tissues, or organisms.

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7 protocols using 2 deoxy d 1 3h glucose

Measuring Glucose and Glutamine Uptake

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Cells (1 × 10⁶) were starved by incubation at 37 °C in serum/glucose-free or glutamine-free RPMI for 30 min. Radiolabelled 2-deoxy-d-[1-³H]glucose or glutamine-l-[3,4-³H(N)] (PerkinElmer) was added to a final concentration of 0.1 mM (2 μCi ml⁻¹). Cells were incubated for 10 min at room temperature, washed in cold serum/glucose/glutamine-free RPMI, and solubilized in 500 μl 0.1% SDS. Radioactivity was measured by liquid scintillation.

Clerc I., Moussa D.A., Vahlas Z., Tardito S., Oburoglu L., Hope T.J., Sitbon M., Dardalhon V., Mongellaz C, & Taylor N. (2019). Entry of glucose- and glutamine-derived carbons into the citric acid cycle supports early steps of HIV-1 infection in CD4 T cells. Nature metabolism, 1(7), 717-730.

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Evaluation of MD-1 Capsule Constituents

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MD-1 capsules (3 different batches) were purchased from Isha Arogya, Chennai, Tamilnadu. The capsule contents were collected from twenty capsules of each batch and stored in a amber colour container at room temperature. Calibration standards for heavy metal analysis (Cd, Pb, As, Hg) were purchased from Merck, Germany. 2-Deoxy- D-[1-3H] glucose was purchased from PerkinElmer, USA. Insulin, dexamethasone (DEX), Iso Butyl Methyl Xanthine (IBMX) were purchased from Sigma Aldrich, USA. Pioglitazone (PGZ) was a kind gift from Dr. Reddys Laboratories, India. All other chemicals and solvents were of analytical grade obtained from SISCO Research Laboratories Pvt Ltd. India.

Telapolu S., Kalachavedu M., Punnoose A.M, & Bilikere D. (2018). MD-1, a poly herbal formulation indicated in diabetes mellitus ameliorates glucose uptake and inhibits adipogenesis – an in vitro study. BMC Complementary and Alternative Medicine, 18, 113.

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Insulin Signaling Pathway Analysis

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Insulin, BSA, anti-β-actin antibody and propranolol hydrochloride were from Sigma-Aldrich (St. Louis, MO, USA). Zinc-Alpha-2-Glycoprotein (ZAG) Human HEK293 recombinant protein was purchased from Biovendor (Laboratory Medicine Inc., Palackeho, Czech Republic). The β1-adrenoceptor antagonist 1-[2-((3-Carbamoyl-4-hydroxy)phenoxy)ethylamino]-3-[4-(1-methyl-4-trifluoromethyl-2-imidazolyl)phenoxy]-2-propanol dihydrochloride (CGP-20712A) was from Tocris Bioscience (Bristol, UK). Culture media and sera were from Invitrogen (Paisley, UK). 2-Deoxy-D-[1-3H] glucose (11.0 Ci/mmol) was from PerkinElmer Life Sciences (Boston, MA, USA). Antibodies against phosphorylated (p)-IRS-1 (Tyr612), total IRS-1 and PP2A were from Millipore (Bedford, MA, USA). Antibodies against p-AKT1 (Ser473), total AKT, p-AS160 (Thr642) and total AS160 were from Cell Signaling Technology (Beverly, MA, USA).

Ceperuelo-Mallafré V., Ejarque M., Duran X., Pachón G., Vázquez-Carballo A., Roche K., Núñez-Roa C., Garrido-Sánchez L., Tinahones F.J., Vendrell J, & Fernández-Veledo S. (2015). Zinc-α2-Glycoprotein Modulates AKT-Dependent Insulin Signaling in Human Adipocytes by Activation of the PP2A Phosphatase. PLoS ONE, 10(6), e0129644.

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Radiolabeled Glucose Uptake Assay

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The measurement of radiolabelled glucose uptake has been described previously (21 (link)). Briefly, glucose-free RMPI 1640 medium labelled with 0.5 µCi/ml radiolabelled 2-deoxy-d-[1-3H]glucose (Perkin Elmer) was layered over 500 µl of a 1:1 mixture of silicone oil (Sigma Aldrich) and dibutyl phthalate (Sigma Aldrich). Cells (1 x 106 per measurement) were added and allowed to take up radiolabelled 2-deoxy-d-[1-3H]glucose for 4 min. The cells were then lysed in 100 mM NaOH and the radioactivity was analyzed using a Beckman LS 6500 Multi-Purpose Scintillation Counter (Beckman Coulter). Assays were performed in duplicate or triplicate per biological replicate. For each biological replicate, data were normalized to the uptake of 2-deoxy-d-[1-3H]glucose in the IL-2-maintained CTL sample.

Rollings C.M., Sinclair L.V., Brady H.J., Cantrell D.A, & Ross S.H. (2018). Interleukin-2 shapes the cytotoxic T cell proteome and immune environment-sensing programs. Science signaling, 11(526), eaap8112.

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2-Deoxy-D-[1-3H]Glucose Uptake Assay

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Cells (1 × 10⁶ cells) were grown in six-well plates and incubated for 24 h. Cells were incubated with 1 μCi (1 μl) of 2-deoxy-d-[1-³H]glucose (Perkin Elmer) in 1 ml of media for 20 min at 37°C. The cells were washed two times with ice-cold PBS, and lysed in 1 ml of 1% SDS. Cell lysate 0.8 ml in 4.5 ml Ecoscint H (National Diagnostics LS-275) was counted on a scintillation device for 1 min. All experiments were done in triplicates wells, and repeated at least two times.

Leung E.Y., Kim J.E., Askarian-Amiri M., Joseph W.R., McKeage M.J, & Baguley B.C. (2014). Hormone Resistance in Two MCF-7 Breast Cancer Cell Lines is Associated with Reduced mTOR Signaling, Decreased Glycolysis, and Increased Sensitivity to Cytotoxic Drugs. Frontiers in Oncology, 4, 221.

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Glucose Uptake Assay for T Cells

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Prior to transport analyses, T cells (0.5 × 10⁶) were starved in glucose-free RPMI for 30 min at 37 °C. Glucose uptake was initiated by addition of 2-deoxy-D[1-³H]glucose (2 μCi; Perkin Elmer) for 10 min at room temperature. Cells were then washed and lysed in 0.1% SDS (500 μl) and radioactivity was measured by liquid scintillation. Uptake for each cell subset is expressed as mean counts per minute (CPM) for triplicate samples and error bars indicate standard deviation (SD).

Cretenet G., Clerc I., Matias M., Loisel S., Craveiro M., Oburoglu L., Kinet S., Mongellaz C., Dardalhon V, & Taylor N. (2016). Cell surface Glut1 levels distinguish human CD4 and CD8 T lymphocyte subsets with distinct effector functions. Scientific Reports, 6, 24129.

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Isoproterenol-Induced Cellular Signaling Assay

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Isoproterenol, insulin, collagenase type II, 8-Br-cAMP, latrunculin B, and DMEM (4.5 g d-glucose/liter) were all obtained from Sigma-Aldrich. Compound 15e (3-[4-(4-morpholinyl)thieno[3,2-d]pyrimidin-2-yl]-phenol), PI-103 (3-(4-(4-morpholinyl), pyrido[3′,2’:4,5] furo [3,2-d]pyrimidin-2-yl)phenol), LY294002, and rapamycin were from Alexis Biochemical. KU 0063794 was obtained from Axon Medchem. Akt inhibitor C was obtained from EMD Millipore, 2-deoxy-d-[1-³H]-glucose ([³H]2DG; specific activity 7.5 Ci/mmol) from PerkinElmer. Torin1 was provided by D.M. Sabatini (Whitehead Institute, Cambridge, MA).

Olsen J.M., Sato M., Dallner O.S., Sandström A.L., Pisani D.F., Chambard J.C., Amri E.Z., Hutchinson D.S, & Bengtsson T. (2014). Glucose uptake in brown fat cells is dependent on mTOR complex 2–promoted GLUT1 translocation. The Journal of Cell Biology, 207(3), 365-374.

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