Subcutaneous and Visceral adipose tissues were processed for metabolite analysis40 (link),88 (link). The samples were quickly snap-frozen with liquid nitrogen and incubated in – 80 °C for 10 min. The tissues were quickly ground and blended in a mortar. Powdered fat tissues (20 mg) were then placed into a 1.5 mL scaled Eppendorf tube containing 300 μl lysis buffer (9:1 methanol: chloroform), mixed with 500 μL water. The samples were immediately vortexed for 3 min, and the tissues were subjected to centrifugation at 16,000×g for 5 min at 4 °C to remove the cell debris. The supernatant was stored at − 80 °C, before proceeding for LC–MS/MS analysis. Glucose-1-Phosphate, Glucose-6-Phosphate, Fructose-1-Phosphate, Fructose-1,6-bisphosphate, Pyruvate, PhosphoenolPyruvate, and the standards were purchased from Sigma-Aldrich, USA. Chloramphenicol (internal standard) (Sigma-Aldrich, USA) was prepared and diluted to 100 ng/mL using methanol.
Glucose 1 phosphate
Manufactured by Merck Group
Sourced in United States
Glucose-1-phosphate is a laboratory reagent used for biochemical and enzymatic analyses. It is a derivative of glucose that plays a crucial role in carbohydrate metabolism. The primary function of Glucose-1-phosphate is to serve as a substrate or intermediate in various metabolic pathways, enabling researchers to study and understand the complex processes involved in glucose utilization and energy production within biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Glucose 6 phosphate (g6p), by Merck Group (4 mentions)
Mgcl2, by Merck Group (3 mentions)
Adenosine triphosphate (atp), by Merck Group (3 mentions)
Acetonitrile, by Merck Group (2 mentions)
Glycine, by Merck Group (2 mentions)
Valine, by Merck Group (2 mentions)
Hepes, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Biomol green, by Enzo Life Sciences (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Adenosine monophosphate, by Merck Group (2 mentions)
Fructose 6 phosphate, by Merck Group (2 mentions)
Adenosine diphosphate (adp), by Merck Group (2 mentions)
Chloramphenicol, by Merck Group (1 mentions)
Pyruvate, by Merck Group (1 mentions)
Phosphoenolpyruvate, by Merck Group (1 mentions)
Fructose 1 6 bisphosphate, by Merck Group (1 mentions)
Reduced glutathione gsh, by Merck Group (1 mentions)
Anthrone, by Merck Group (1 mentions)
1 chloro 2 4 dinitrobenzene, by Merck Group (1 mentions)
11 protocols using glucose 1 phosphate
1
Adipose Tissue Metabolite Profiling
2
Streptozotocin-Induced Diabetes Assays
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NA, STZ [2-deoxy-2-(3-methyl-3nitrosoureido)-D-glycopyranoside], glucose-6-phosphate, glucose-1-phosphate, anthrone, reduced glutathione (GSH), malondialdehyde (MDA), and 1-Chloro-2,4-dinitrobenzene were purchased from Sigma-Aldrich Chemical Co., St Louis, MO, USA. All other chemicals were of analytical grade and were obtained from standard commercial supplies.
3
HPLC-based Metabolite Quantification
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HPLC-grade methanol and acetonitrile were purchased from Merck Chemicals (Darmstadt, Germany). BSTFA (containing 1% TMCS) was purchased from REGIS Technologies (Morton Grove, IL, USA). Epicatechin, citrulline, glucose-1-phosphate, fructose, glutamine, glycine, alanine, proline, serine, tryptophan and valine were purchased from Sigma-Aldrich (St. Louis, MO, USA).
4
Enzymatic Kinetic Assay Protocol
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Hepes, glucose 1-phosphate, AMP were purchased from “Sigma” (USA), NaCl was purchased from “Reakhim” (Russia), glycogen was purchased from “Olaine” (Latvia). All solutions for the experiments were prepared using deionized water obtained with the Easy-Pure II RF system (Barnstead, USA).
5
Glycogen Phosphorylase Activity Assay
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The following reagents were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA): dimethylsulfoxide (DMSO), caffeine, CP-91149, ellagic acid, HEPES, MgCl2, glucose 1-phosphate, and glycogen. KCl was purchased from José M. Vaz Pereira, S.A. (Sintra, Lisboa). Rabbit muscle GPa was purchased from Creative Enzymes® (Shirley, NY, USA). The reagent for colorimetric phosphate quantitation, BIOMOL® Green, was acquired from Enzo Life Sciences, Inc. (Miraflores, Portugal).
6
Enzymatic Assay for Melatonin Synthesis
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AIA was a gift from Roche Co. (Germany). DDC was purchased from Aldrich Chemical Company Inc. (Milwaukee, WI). Melatonin, bovine serum albumin, deoxyguanosine 5′-diphosphate, glucose-1-phosphate, malate dehydrogenase, NADH, Dowex 1, Dowex 50 W and thiobarbituric acid were obtained from Sigma Chemical Co. (St. Louis, MO). All other chemicals were of analytical grade.
7
Metabolite Standard Preparation Protocol
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Pure metabolites threonine, valine, isoleucine, glutamic acid, adenosine monophosphate (AMP), adenosine triphosphate (ATP), 3-phosphoglyceric acid, glucose-1-phosphate, fructose-6-phosphate, fructose-1,6-biphosphate, itaconic acid, succinic acid and citric acid were obtained from Sigma-Aldrich (St. Louis, USA). Stock individual standard solutions (500 μg mL−1) were prepared dissolving accurate amounts of pure standards in Acetonitrile:water 1:1. Two standard mixture samples of these compounds were prepared at 10 and 20 μg mL−1 concentration levels in acentonitrile:water 1:1. Ethanol, Acetonitrile and HPLC grade water were obtained from Merck (Darmstadt, Germany).
8
Enzymatic Biosynthesis of Nucleotide Sugars
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DNaseI, Glucose-1-phosphate, dTTP, dTDP-D-glucose, malachite green reagent, NADPH, phenylmethanesulfonyl fluoride (PMSF), pyrophosphatase, UTP and UDP-glucose were obtained from Sigma-Aldrich (St. Louis MO), dTDP-4-keto-6-deoxy-glucose came from Carbosynth (Berkshire, UK), novobiocin and ampicillin were obtained from Duchefa Biochemie (Haarlem, The Netherlands), while restriction endonucleases were purchased from Promega (Madison, WI).
9
Biomolecular Characterization of Cell Signaling
10
Synthesis and Evaluation of Glycogen Phosphorylase Inhibitors
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The following reagents were obtained from Sigma-Aldrich, Inc. (St. Louis, MO): dimethylsulfoxide (DMSO), 5-chloro-N-[(1S,2R)-3-(dimethylamino)-2-hydroxy-3-oxo-1-(phenylmethyl)propyl]-1H-indole-2-carboxamide (CP-91149 ), HEPES, MgCl2, glucose 1-phosphate, glycogen, and compounds 42 , 45 and 47 . KCl was obtained from José M. Vaz Pereira, S.A. (Sintra, Lisboa). The rabbit muscle phosphorylated form, mGPa, was obtained from Creative Enzymes® (Shirley, NY, USA). The reagent for colorimetric phosphate quantitation, BIOMOL® Green, was obtained from Enzo Life Sciences, Inc. (Postfach, Lausen, Switzerland). Compounds 36 –41 and 44 were obtained from INDOFINE Chemical Company, Inc. (Hillsborough, NJ, USA). Compounds 46 and 48 –52 were obtained from Extrasynthese (Lyon Nord, Genay, France). Compound 43 was obtained from Alfa Aesar (Erlenbachweg, Kandel, Germany). Compounds 1 –35 were synthesized as previously described [27 (link),29 (link),30 (link),31 (link),32 (link),33 (link),34 (link),35 (link)].
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