Methanol and dichloromethane were purchased from Scharlau SL (Sentmenat, Spain), deuterium oxide (D2O) and sodium 3-trimethylsilyl-2,2,3,3-tetradeuteriopropionate (TSP) from Eurisotop (Saint-Aubin, France). Hippuric acid, 3-hydroxybutyric acid, citric acid, formic acid, and L-Alanine were purchased from Sigma Chemical Company (Saint Louis, MO).
3 hydroxybutyric acid
Manufactured by Merck Group
Sourced in United States
3-hydroxybutyric acid is a chemical compound that can be used in laboratory settings. It is a type of organic acid that is naturally present in the human body. This compound may have various applications in research and analysis, but no further details about its specific uses are provided.
Automatically generated - may contain errors
Lab products found in correlation
L alanine, by Merck Group (3 mentions)
Malic acid, by Merck Group (3 mentions)
Fumaric acid, by Merck Group (3 mentions)
Succinic acid, by Merck Group (3 mentions)
Formic acid, by Merck Group (2 mentions)
Citric acid, by Merck Group (2 mentions)
L valine, by Merck Group (2 mentions)
L isoleucine, by Merck Group (2 mentions)
L serine, by Merck Group (2 mentions)
Glycine, by Merck Group (2 mentions)
L leucine, by Merck Group (2 mentions)
L glutamic acid, by Merck Group (2 mentions)
α ketoglutaric acid, by Merck Group (2 mentions)
Deuterium oxide d2o, by Eurisotop (1 mentions)
Hippuric acid, by Merck Group (1 mentions)
Urea, by Thermo Fisher Scientific (1 mentions)
Palmitic acid, by Thermo Fisher Scientific (1 mentions)
Taurocholic acid, by Steraloids (1 mentions)
Sodium taurochenodeoxycholate, by Merck Group (1 mentions)
Serotonin, by Merck Group (1 mentions)
9 protocols using 3 hydroxybutyric acid
1
Metabolite Extraction and Identification
2
Comprehensive Metabolite Profiling Protocol
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Uric acid, L-tryptophan, L-alanine, L-valine, L-leucine, L-isoleucine, glycine, l-serine, succinic acid, fumaric acid, malic acid, 2-ketoglutaric acid, L-glutamic acid, D-ribose, sodium 2-hydroxybutyrate, 3-hydroxybutyric acid, 2-isopropylmalic acid, formic acid, serotonin, γ-aminobutyric acid-2,2,3,3,4,4d6, 21-deoxycortisol, 11-deoxycortisol, oleic acid, cis-4,7,10,13,16,19-docosahexaenoic acid, cis-10-nonadecenoic acid (C19:1n9c) and sodium taurochenodeoxycholate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Taurocholic acid was obtained from Steraloids (Newport, RI, USA). p-Cresyl sulfate potassium salt and arachidonic acid were supplied from Toronto Research Chemicals (North York, ON, Canada). Urea, HPLC-graded acetonitrile, methanol and isopropanol were obtained from Fisher Scientific (Hampton, NH, USA). Water was purified in-house using a Milli-Q Advantage A10 water purification system (Millipore, Bedford, MA, USA). Palmitic acid, cis-9,12-linoleic acid, 2-aminobutyric acid, pyridine (HPLC grade) with AcroSeal and N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) were acquired from Acros Organics (Morris Plains, NJ, USA).
3
Transfection and Luciferase Assay in 3T3-L1 Cells
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On day 7 after the induction of differentiation as stated above, the medium of 3T3-L1 cells in 12-well plates was changed to Opti-MEM (Invitrogen, Carlsbad, CA), and the cells were transfected with luciferase reporter plasmids using Lipofectamine 3000 Transfection Reagent (Invitrogen) according to the protocol provided by the manufacturer. Transfection was performed using 2 ng CMV-Renilla (internal standard) and 100 ng reporter plasmids. The next day (day 8), the medium was changed to Krebs-Ringer Bicarbonate buffer, composed as stated above, with or without 10 mM 3-Hydroxybutyric acid (Sigma) or 1 μM pioglitazone (Takeda, Osaka, Japan). After 24-hr incubation, luciferase reporter assays were performed using Promega Dual-Luciferase Reporter Assay System (Promega). Luciferase values were normalized by an internal CMV-Renilla control and expressed as relative luciferase activity.
4
Inducing Adipogenesis in 3T3-L1 Cells
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On day 7 after differentiation, the medium of 3T3-L1 adipocytes was replaced with DMEM composed of 2.5 mM glucose with 10 mM concentrations of 3-hydroxybutyric acid (Sigma–Aldrich). After 24 h (day 8), 3T3-L1 adipocytes were additively stimulated by 1 nM insulin for 24 h, followed by harvesting (day 9).
5
Comprehensive Chemical Characterization
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Isopropyl alcohol (IPA), methanol (MeOH), and ethanol (EtOH) (all LC–MS-grade) were purchased from Fisher Scientific (UK). N,N-Dimethylformamide (DMF) and chloroform (CHCl3) were purchased from EMPLURA Merck Millipore (BE) and Sigma-Aldrich (US), respectively. Ultrapure water (UPW) was obtained through a Sartorius Arium 661 UV water purification system (Millipore, Belgium). Analytical standards [3-hydroxybutyric acid, linoleic acid, l -carnosine, 1,2-dioleoyl-sn-glycero−3-phospho-rac-(1-glycerol) (DOPG), 1-palmitoyl−2-oleoyl-sn-glycero−3-(phospho-rac-(1-glycerol)) (POPG), D-mannitol, myo-inositol, l -cysteine, l -arginine, l -kynurenine, and taurolitocholate-3-sulfate], and leucine enkephalin were purchased from Sigma-Aldrich (US), ICN Biomedicals Inc. (US), TLC Pharmchem (CA), or Waters Corporation (US) or as reported in (11 (link), 77 (link)). Information on the purchased polymers and initial evaluation of different polymer solutions is given under note S1.
6
Adipocyte Differentiation Metabolism Assay
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On day 7 after the induction of differentiation, the medium of 3T3-L1 cells was replaced with Krebs-Ringer Bicarbonate buffer (KRBB), composed of 25 mM NaHCO3 (Nacalai), 119 mM NaCl (Nacalai), 4.74 mM KCl (Nacalai), 1.19 mM MgCl2 (Nacalai), 1.19 mM KH2PO4 (Nacalai), 2.54 mM CaCl2 (Nacalai), 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Nacalai), 0.05 mM Bovine Serum Albumin solution (Sigma), and 25 mM glucose (Otsuka), with various concentrations of 3-Hydroxybutyric acid (Sigma). The cells were collected after 24 hr, then grown at 37 °C under 5% CO2 fully humidified air environment.
7
Metabolic Profiling Using GC-MS
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The following OA standards used in this study were purchased from Sigma-Aldrich (St. Louis, MO, USA): 3,4-dimethoxybenzoic acid as internal standard (IS), 3-hydroxybutyric acid, pyruvic acid, lactic acid, succinic acid, fumaric acid, oxaloacetic acid, α-ketoglutaric acid, malic acid, cis-aconitic acid, citric acid and isocitric acid. Acetoacetic acid was purchased from Tokyo Chemical Industry (Tokyo, Japan). Methoxyamine hydrochloride was also obtained from Sigma-Aldrich. N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA)+1% tert-butyldimethylchlorosilane was obtained from Thermo Scientific (Bellefonte, PA, USA). Toluene, diethyl ether, ethyl acetate and dichloromethane (pesticide grade) were purchased from Kanto Chemical (Chuo-ku, Tokyo, Japan). Sodium hydroxide was supplied by Duksan (Ansan, South Korea), and sulphuric acid was purchased from Samchun Pure Chemical (Pyeongtaek, South Korea). All other chemicals were of analytical reagent grade.
8
Synthesis and Characterization of PHB
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1-Undecene and 1-tridecene were purchased from Alfa Aesar (MA, USA). PHB, benzoic acid, 1-heptene, 1-nonene, benzoic acid and 3-hydroxybutyric acid were purchased from Sigma-Aldrich (MO, USA). Methyl ester of 3-hydroxyoctanoic acid (3-OH C8), 3-hydroxydecanoic acid (3-OH C10), 3-hydroxydodecanoic acid (3-OH C12) and 3-hydoxytetradecanoic acid (3-OH C14) were purchased from Matreya LLC (PA, USA). All solvents and reagents were either of HPLC grade or analytical reagent grade.
9
Analytical Standards for Metabolomic Analysis
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The reference compounds (sodium pyruvate, sodium lactate, citric acid monohydrate, succinic acid, malic acid, fumaric acid, 2-hydroxybutyric acid, oxalic acid, 3-hydroxybutyric acid, glycine, l -serine, l -valine, l -threonine, l -alanine, l -aspartic acid, l -glutamine, l -glutamic acid, l -leucine, l -isoleucine, l -tyrosine, erythronate, meso-erythritol, α-ketoglutaric acid, glyceric acid) were purchased from Sigma-Aldrich, Munich, Germany. U13C-Ribitol (CIL Inc., Tewksbury, MA, USA), pentanedioic-d6-acid (C/D/N isotopes Inc., Quebec, QC, Canada), and norleucine (Sigma-Aldrich, Munich, Germany) were used as internal standards during metabolite extraction. U13C-Glutamate, U13C-glucose, and U13C-glutamine were purchased from CIL Inc., Tewksbury, MA, USA. For metabolite extraction, high purity HPLC (or higher) grade solvents methanol, acetone, isopropanol, and acetonitrile, and MQ water (18.2 M·cm, <3 ppb TOC) were used.
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