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3 protocols using potassium hydroxycitrate tribasic monohydrate

1

Synthesis of Citrate Compounds

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The following reagents were purchased from Sigma–Aldrich (St. Louis, MO): melamine, d,l-isocitrate acid trisodium salt hydrate (93%), potassium hydroxycitrate tribasic monohydrate (95%), potassium citrate (Kcit), and etidronic acid (EtA). All reagents were used as received without further purification.
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2

Evaluation of Garcinia cambogia Compounds

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Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), penicillin/streptomycin, and trypsin-ethylenediaminetetraacetic acid (EDTA) were obtained from Gibco, Inc. (Thermo Fisher Scientific, Waltham, MA, USA). Sodium oleate, sodium palmitate, oil red O, isopropanol, (−)-epigallocatechin gallate (EGCG), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), digitonin, N-acetyl-L-cysteine (NAC), anti-rabbit-fluorescein isothiocyanate (FITC, F0382), and potassium hydroxycitrate tribasic monohydrate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin was obtained from Bovogen Biologicals (Melbourne, Australia). Anti-C/EBPα (8178), anti-PPARγ (2443), anti-BAX (2772), anti-PARP (9532), anti-NRF2 (12721), anti-Lamin A/C (4777), and goat anti-rabbit (7074) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-Bcl-2 (sc-7382) antibody was obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Anti-β-actin (LF-PA0207) and goat anti-mouse (LF-SA8001) antibodies were purchased from Abfrontier (Geumcheon, Seoul, Korea). Garcinia cambogia (G. cambogia) powder (main component: hydroxycitric acid, 59.55%) was obtained from Mirae Biotech, Inc. (Pocheon, Gyeonggi-do, Korea). All other chemicals were of analytical grade.
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3

Aqueous Mineral Compound Synthesis

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The following reagents were used: glutaric acid (C5H8O4, ≥99.0%, H2(Glr), Sigma-Aldrich), potassium hydroxycitrate tribasic monohydrate (K3(C6H5O8)·H2O, ≥ 95.0%, (K3(CitOH), Sigma-Aldrich), sodium citrate tribasic dihydrate (Na3(C6H5O7)·2H2O, ≥ 99.0%, (Na3(Cit), Sigma-Aldrich), sodium hexametaphosphate ((NaPO3)6, ≥96.0%, Sigma-Aldrich), sodium hydroxide (NaOH, ≥ 99.0%, Sigma-Aldrich), sodium phosphate dibasic dihydrate (Na2HPO4·2H2O, ≥ 99.0%, Sigma-Aldrich), and calcium chloride dihydrate (CaCl2·2H2O, ≥ 99.0%, Sigma-Aldrich). All the solutions were made with ultrapure water (Arium pro, Sartorius).
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