Acetonitrile, ethyl acetate, formic acid and methanol were purchased from Kefo (Sisak, Croatia); Czapek Yeast Agar (CYA) was purchased from Oxoid (Hampshire, UK). Medium RPMI 1640, fetal bovine serum (FBS), phosphate buffered saline (PBS; without Ca2+ and Mg2+), trypsin-EDTA, antibiotics penicillin and streptomycin were from Lonza (Basel, Switzerland). The MTT reagent [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], acridine orange, ethidium bromide, normal melting point agarose (NMP), low melting point agarose (LMP), phorbol 12-myristate 13-acetate (PMA), Triton X-100 and Tris buffer were purchased from Sigma-Aldrich (Deisenhofen, Germany) and molecular biology grade dimethyl sulfoxide (DMSO), used for dissolving the extracts and for the control treatment, was purchased from Sigma-Aldrich (Deisenhofen, Germany); technical grade DMSO, used for dissolving the formazane, ethanol, sodium chloride, disodium-EDTA and sodium hydroxide were from Kemika (Zagreb, Croatia).
Ethanol
Manufactured by Kemika
Sourced in Croatia
Ethanol is a clear, colorless liquid used as a solvent and chemical intermediate in various laboratory applications. It has a specific gravity of approximately 0.789 and a boiling point of 78.3°C. Ethanol is a volatile organic compound that is miscible with water and many other organic solvents.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Kemika (2 mentions)
Normal melting point agarose, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Kemika (1 mentions)
Ethidium bromide, by Merck Group (1 mentions)
Acridine orange, by Merck Group (1 mentions)
Tris buffer, by Merck Group (1 mentions)
Czapek yeast agar, by Thermo Fisher Scientific (1 mentions)
Low melting point agarose lmp, by Merck Group (1 mentions)
Molecular biology grade dimethyl sulfoxide dmso, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Mtt reagent, by Merck Group (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Potassium chloride (kcl), by Kemika (1 mentions)
Cacl2, by Kemika (1 mentions)
Nahco3, by Kemika (1 mentions)
Pepsin, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Myricetin, by Extrasynthese (1 mentions)
6 protocols using ethanol
1
Cytotoxicity Assay Protocol
2
Analytical Reagents for Research Protocols
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All reagents and solvents used for the analyses were of analytical grade. Pepsin was obtained from Sigma Aldrich GmbH (Taufkir-chen, Germany), salts (NaCl, KCl, NaH2PO4, Na2SO4, NaHCO3, CO(NH2)2, CaCl2), ethanol, and hydrochloride acid were obtained from Kemika (Zagreb, Croatia). DPPH was obtained from Merck KgaA (Darmstadt, Germany).
3
HPLC Analysis of Phenolic Compounds
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All chemicals used were of high-performance liquid chromatography (HPLC) analytical grade. Acetonitrile was purchased from from J.T. Baker (Deventer, The Netherlands). Formic acid and orthophosphoric acid (85% w/w) were purchased from Fluka (Buchs, Switzerland) and ethanol from Kemika (Zagreb, Croatia). Phenolic standard compounds of analytical standard grade used for identification and quantification were: delphinidin-3-O-glucoside, cyanidin-3-O-glucoside, peonidin-3-O-glucoside, malvidin-3-O-glucoside, epigallocatechin, procyanidin B1, procyanidin B2, rutin (quercetin 3-O-rutinoside) and myricetin from Extrasynthese (Lyon, France); (–)-epicatechin, (+)-catechin and (–)-and epicatechin-gallate from Sigma-Aldrich (St. Louis, MO, USA); kaempferol, quercetin and isorhamnetin from Fluka (Steinheim, Germany) and quercetin-3-O-glucoside from Sigma (St. Louis, MO, USA).
4
Analyzing Antioxidant and Lipid Properties of Propolis
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All chemicals to investigate the lipidemic properties, the antioxidant capacity, and the analytics of propolis were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA) except ethanol, methanol, potassium acetate, potassium hydroxide, and sodium carbonate, which were purchased from Kemika (Zagreb, Croatia) or from Merck, Darmstadt, Germany (AlCl3 and 2,4-dinitrophenylydrazine). Perkin Elmer Lambda 25 spectrophotometer (Perkin Elmer, Waltham, MA, USA) was used for the absorbance measurements.
5
Cytotoxicity of Rare Honey Varieties
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We ran this assay, as this type of honey is quite rare and its safety profile poorly known. In vitro cytotoxicity was determined as described by Babich and Borenfreund (19 (link)). Briefly, CAL 27, HepG2, and Caco-2 cells were seeded in 96-well plates (105 cells per mL) and treated with STH, STHE, HGA, and AFH samples in four concentrations (Table 1 ) for 30 min, 1 h, and 2 h. After treatment, we added Neutral Red (Sigma-Aldrich) solution, incubated the cells at 37 °C for 1 h, removed Neutral Red solution, and washed the cells with phosphate buffered saline (PBS) (100 μL). Finally, we added 100 μL of solution combining acetic acid (Sigma-Aldrich), ethanol (Kemika), and demineralised water in the 0.1:5:4.9 ratio to extract the dye from the lysosomes of living cells. Cell viability was measured as colour intensity and absorption at 540 nm in a FLUOstar OPTIMA plate reader (BMG Labtech, Durham, NC, USA).
Data are expressed as percentage of viability of untreated cells (considered to be 100 %). Each concentration was tested in quadruplicate, and each experiment was repeated three times.
Data are expressed as percentage of viability of untreated cells (considered to be 100 %). Each concentration was tested in quadruplicate, and each experiment was repeated three times.
6
Surface Free Energy Determination of Alloys
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The experiment was performed using an OCA 20 goniometer (Data Physics Instruments, Filderstadt, Germany). The contact angle was determined by the sessile drop method. The instrument was equipped with an automatic drop dosing system and software for image analysis and determination of the contact angle between the test liquid droplet and test material. The test liquids were water and diiodomethane (99+%, Acros Organics, Geel, Belgium). Prior to the measurements, the wire surface was cleaned with 95% ethanol (Kemika, Zagreb, Croatia) and allowed to dry. Then, 0.3 μL of the test liquid was applied. Ten measurements per alloy type were performed at a consistent temperature of 25°C. The dispersive and polar components and total SFE of the probe liquids used for the SFE calculation of alloys were as follows: 21.8 (dispersive), 51.0 (polar), and 72.8 (total) for water and 50.8, 0.0 and 50.8 mJm -2 for diiodomethane. The free surface energy was determined indirectly by measuring the contact angles of the two liquids and calculated using mathematical models by Owens-Wendt and Wu 28, 29) .
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