Licochalcone A (1 ) and demethylzeylasteral (DZ) were purchased from Biopurify Phytochemicals Ltd. (Chengdu, China) and confirmed by 1H-NMR. Licochalcones B, C, H and xanthohumol (2 –5 ) were prepared using previously reported methods [27 (link),28 (link)]. All the microorganisms were obtained from the Korean Collection for Type Cultures (KCTC). All of the ingredients for microbial media including dextrose, peptone, malt extract, yeast extract, and potato dextrose broth were purchased from Becton, Dickinson and Company (Sparks, MD, USA). Dulbecco’s Modified Eagle Medium (DMEM) and Antibiotic-Antimycotic were purchased from Gibco (Invitrogen, Carlsbad, CA, USA); fetal bovine serum (FBS) was purchased from Welgene Inc. (Gyeongsan-si, Korea). Phosphate-buffered saline (PBS) tablets were purchased from Takara Korea Biomedical Inc. (Seoul, Korea) and thiazolyl blue tetrazolium bromide (MTT) was from Thermo Fisher Scientific (Waltham, MA, USA).
Thiazolyl blue tetrazolium bromide mtt
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
Thiazolyl Blue tetrazolium bromide (MTT) is a colorimetric assay used to measure cell metabolic activity. It is a yellow tetrazolium salt that is reduced by metabolically active cells to produce a purple formazan dye, which can be quantified spectrophotometrically.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Formalin, by Merck Group (2 mentions)
Isopropanol, by Merck Group (2 mentions)
Insulin, by Merck Group (2 mentions)
Oil red o, by Merck Group (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Raw264.7 cell, by American Type Culture Collection (2 mentions)
Griess reagent, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Welgene (1 mentions)
Dextrose, by BD (1 mentions)
Malt extract, by BD (1 mentions)
Peptone, by BD (1 mentions)
Yeast extract, by BD (1 mentions)
Potato dextrose broth (pdb), by BD (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Tetraethyl orthosilicate teos, by Thermo Fisher Scientific (1 mentions)
Acetic acid hoac, by Merck Group (1 mentions)
Ammonium hydroxide nh4oh, by Thermo Fisher Scientific (1 mentions)
Blue dextran, by Merck Group (1 mentions)
14 protocols using thiazolyl blue tetrazolium bromide mtt
1
Isolation and Evaluation of Licochalcones
2
Synthesis and Characterization of Hyaluronic Acid Hydrogels
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Tetraethyl orthosilicate (TEOS) was purchased from Acros Organics (TEOS A, Geel, Belgium) and Alfa Aesar (TEOS B, Lancashire, UK), both now Thermo Scientific (Waltham, MA, USA). Sodium Hyaluronan (HA, 5 kDa) was from Lifecore Biomedical (HA A, Chaska, MN, USA) and HTL Biomedical (HA B, Javené, France). Acetic acid (HOAc) was from EMD Millipore (Billerica, MA, USA), ammonium hydroxide (NH4OH) was from Fisher Chemical (Fair Lawn, NJ, USA), and phosphate-buffered saline (PBS) was from Corning Life Sciences (Durham, NC, USA). Fluorescein was from Fluka Analytical, owned by Sigma Aldrich (St. Louis, MO, USA), fluorescein disodium was from Alfa Aesar, now Thermo Scientific (Waltham, MA, USA), green fluorescent protein (GFP) was from Novus Biologicals (Littleton, CO, USA), and blue dextran with molecular weights of 5597, 15,038, and 500,000 g/mol, referred to as 5000, 20,000, and 500,000 g/mol, respectively, was from Sigma Aldrich (St. Louis, MO, USA). Skin Irritation Test kit (SPI-200-SIT) was from MatTek (Ashland, MA, USA), thiazolyl blue tetrazolium bromide (MTT) was from Thermo Scientific (Waltham, MA, USA), and isopropanol was from VWR (Radnor, PA, USA). Hyaluronidase was from MP Biomedicals (Solon, OH, USA), and hydrochloric acid (HCl) was from Janssen Pharmaceuticals (Beerse, Belgium).
3
Anti-adipogenic Assays with Mouse Fibroblasts
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Mouse embryonic fibroblast (3T3-L1 preadipocyte) supplied by American type culture collection (ATCC) was used for anti-adipogenic assays. Dulbecco’s modified Eagle medium (DMEM), newborn calf serum (NCS) and fetal bovine serum (FBS) (Life Technologies Corporation, Waltham, MA, USA), 3-isobutyl-1methylxanthine (IBMX), dexamethasone (DXM), insulin, 10% formalin, isopropanol and Oil Red O (ORO; Sigma-Aldrich, Darmstadt, Germany), thiazolyl blue tetrazolium bromide (MTT; Alfa Aesar, Heysham, UK), lysis reagent (Quiagen Sciences, Germantown, MD, USA), Power Cyber Green and high capacity RNA-cDNA kit (Applied Biosystem, Warrington, UK), RIPA buffer (Thermo Scientific, Bedford, MA, USA), primary antibodies (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and secondary antibodies (Millipore, Burlington, MA, USA) were used for cell assays.
4
Quantifying Biofilm Susceptibility to Ciprofloxacin and LxA4
5
Cytotoxicity Assay of Metal-based Compounds
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Copper(II) chloride dehydrate (CuCl2·2H2O), hydrochloric acid (HCl, 37%), nitric acid (HNO3, 65%), and Dulbecco’s modified Eagle’s medium-high glucose (DMEM/HG) were purchased from Thermo Fisher Scientific (Massachusetts, USA). Iron(II) chloride anhydrous (FeCl2, 99.5%), methylene blue (C16H18ClN3) and thiazolyl blue tetrazolium bromide (MTT) were from Alfa Aesar (Massachusetts, USA). Hydrazine hydrate (N2H4·H2O) and sodium pyruvate (C3H3NaO3S·xH2O) were from Acros Organics (Morris Plains, NJ, USA). poly(styrene-alt-maleic acid) sodium salt solution (PSMA), hydrogen peroxide (H2O2), 2′,7′-dichlorofluorescein diacetate (DCFH-DA), sodium bicarbonate (NaHCO3), trypan blue (0.4%), imidazole, N,N-dimethyl-4-nitrosoaniline (RNO), N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), and folic acid (C19H19N7O6) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum (FBS), trypsin-ethylenediaminetetraacetic acid(trypsin-EDTA, 0.25%) and antibiotic-antimycotic (penicillin/streptomycin/amphotericin B) were from Biological Industries (Cromwell, CT, USA). Dimethyl sulfoxide was from Scharlau (Barcelona, Spain).
6
Synthesis and Characterization of CFL Lipid Probes
7
Cell Viability and Adipogenesis Assays
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Solvents including ethanol, methanol, ethyl acetate, hexane, n-butanol, dichloromethane were purchased from SK Chemicals Co. Ltd. (Seongnam, Korea). Dimethyl sulfoxide (DMSO) was purchased from Junsei Chemicals Co. Ltd. (Tokyo, Japan), and sulfanilic acid, N-(1-napthyl) ethylenediamine dihydrochloride, was purchased from Sigma-Aldrich (MO, USA). Cell experiments: RAW 264.7 macrophage cells and 3T3-L1 preadipocytes cells were obtained from the American Type Culture Collection (Rockville, MD, USA). Dulbecco’s modified eagle medium (DMEM), newborn calf serum (NCS), and fetal bovine serum (FBS) were obtained from Gibco-Thermo-Fisher Scientific (Grand Island, NY, USA). 3-Isobutyl-1-methylxanthine (IBMX), dexamethasone, insulin, 10% formalin, isopropanol, and oil red O (ORO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Thiazolyl blue tetrazolium bromide (MTT), used to assess cell viability, was purchased from Alfa Aesar (Tewksbury, MA, USA).
8
Antioxidant and Anti-Inflammatory Assays
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Aluminum chloride (AlCl3) and Folin–Ciocalteu phenol reagent were purchased from Fisher Scientific, India. Ascorbic acid was purchased from Qualigens Fine Chemicals, India. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was from Wako Pure Chemicals, Japan. Ferric chloride (FeCl3) was from Rasayan Laboratories, India. 3T3-L1 preadipocyte and RAW 264.7 cells were obtained from the American Type Culture Collection (ATCC). Bacteria were isolated from clinical samples, identified and supplied by Manipal Teaching Hospital, Phulbari Pokhara. Growth medium (Dulbecco’s modified Eagle’s medium [DMEM]), penicillin and streptomycin antibiotic solution, phenol red-free DMEM, fetal bovine serum (FBS), and newborn calf serum (NCS) were from Life Technologies Corporation, Waltham, MA, USA. Components of the differentiation media (3-isobutyl-1methylxanthine [IBMX], dexamethasone [DXM], and insulin) were from Sigma-Aldrich, Darmstadt, Germany. Isopropanol, 10% formalin, and Oil Red O (ORO) were from Sigma-Aldrich, Darmstadt, Germany. The cell viability assay kit (thiazolyl blue tetrazolium bromide [MTT]) was from Alfa Aesar, Heysham, UK. Dimethyl sulfoxide (DMSO) was from Junsei Chemicals Co Ltd, Japan. Lipopolysaccharide (LPS) from Escherichia coli 0111:B4 and Griess reagent were obtained from Sigma-Aldrich, Darmstadt, Germany.
9
In Vitro Cell Analysis Reagents
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Tyrosine ( purity: ≥98%), tyrosol (≥98%), hydroxytyrosol (≥98%), serotonin (≥98%) and protocatechuic acid (≥97%), fetal bovine serum (FBS), Rosewell Park Memorial Institute (RPMI) 1640 medium, non-essential amino acids (NEAA), minimal essential medium (MEM) vitamins, sodium pyruvate, L-glutamine, antibiotic-antimycotic solution, paraformaldehyde (PFA), bovine serum albumin (BSA) and universal mounting medium (DPX), Ringer HEPES solutions and sodium fluorescein were purchased from Sigma Aldrich (St Louis, MO, USA). Thiazolyl blue tetrazolium bromide (MTT) was obtained from Alfa Aesar (Haverhill, MA, USA). Dimethyl sulfoxide (DMSO), isopropanol, hydrochloric acid, and acetonitrile (ACN) were acquired from PanReac AppliChem (Barcelona, Spain). NuSerum IV was provided by Corning Costar Corp (New York, NY, USA). Triton X-100 was purchased from VWR International (Radnor, PA, USA). β-Catenin (rabbit, #71-2700), Alexa Fluor® 555 (goat anti-rabbit, #A21428), and Hoechst 33342 were acquired from Thermo Fisher Scientific (Waltham, MA, USA).
10
Anti-Inflammatory Effects of E. cava Flake
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Crude E. cava flake (CEF) was obtained from MilaeML (St. Ogeum, Seoul, Korea). RAW264.7 cells were purchased from American Type Culture Collection (Manassas, VA, USA). Dulbecco’s Modified Eagle’s medium (DMEM), Fetal bovine serum (FBS), 100 units/mL penicillin, and 100 μg/mL streptomycin were purchased from Gibco (Grand Island, NY, USA). LPS (Escherichia coli 0111:B4) was purchased from Sigma (St. Louis, MO, USA). Phosphate-buffered saline (PBS) was purchased from RMS Bio-solution (Seoul, Korea). Thiazolyl Blue tetrazolium bromide (MTT) was purchased from Alfa Aesar (Ward Hill, MA, USA). Griess reagent was purchased from Sigma (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits were purchased from BioLegend (San Diego, CA, USA). TRIzol reagent was purchased from Invitrogen (Carlsbad, CA, USA). PD098059 were purchased from Cayman Chemical (Ann Arbor, MI, USA). Primers were purchased from Bioneer (Daejeon, Korea). Antibodies were purchased from Cell Signaling Technology (Boston, MA, USA) and Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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