Intracellular lipids were extracted from samples by either addition of 160 μL ice-cold MeOH to each sample in a microcentrifuge tube or, for samples harvested by filtration, addition of 160 μL ice-cold MeOH to the surface of the cell strainer (held in a microcentrifuge tube, creating an airtight seal which prevents the solvent from passing through the filter). The solvent and microtissues in the microcentrifuge tube or cell strainer were then aspirated and transferred to a 1.75 mL glass vial. An amount of 80 μL ice-cold CHCl3 (HPLC grade, Merck) was added, for a final solvent ratio of 2:1 (v/v) MeOH/CHCl3. For the experiment described in Section 2.3 , MeOH was supplemented with 0.2 μM dodecylphosphorylcholine-d38 (Merck), added as an internal standard to assess technical error resulting from sample preparation and analysis. Samples were vortexed for 2 min and then centrifuged (1500× g, 4 °C) for 20 min. Either 200 μL of the supernatant was transferred to a 1.75 mL glass vial (experiments discussed in Section 2.1 ) or 100 μL of the supernatant was transferred to 200 μL conical glass inserts (experiments described in Section 2.2 and Section 2.3 ). Extracts were then dried by nitrogen blowdown for 10 min. Dried extracts were stored at −80 °C prior to analysis.
Chcl3
Manufactured by Merck Group
Sourced in United States, Germany
CHCl3 is a laboratory chemical compound commonly known as chloroform. It is a colorless, dense liquid with a sweet, ethereal odor. CHCl3 is commonly used as a solvent in various laboratory and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Merck Group (8 mentions)
Hydrochloric acid (hcl), by Merck Group (6 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (6 mentions)
Phosphate buffered saline (pbs), by Lonza (6 mentions)
Sodium hydroxide (naoh), by Merck Group (5 mentions)
Isopropanol, by Merck Group (5 mentions)
Fecl3, by Merck Group (4 mentions)
N n dimethylformamide (dmf), by Merck Group (4 mentions)
N hexane, by Merck Group (4 mentions)
Ch3oh, by Merck Group (4 mentions)
Ascorbic acid, by Merck Group (3 mentions)
Cdcl3, by Merck Group (3 mentions)
Potassium chloride (kcl), by Merck Group (3 mentions)
Boric acid, by Merck Group (3 mentions)
Ch2cl2, by Merck Group (3 mentions)
Alcl3, by Merck Group (2 mentions)
Quercetin, by Merck Group (2 mentions)
Na2co3, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Dicyclohexylcarbodiimide, by Merck Group (2 mentions)
68 protocols using chcl3
1
Intracellular Lipid Extraction Protocol
2
PEOT/PBT and PCL Polymer Solutions
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A polyethylene
terephthalate/polybutylene terephthalate (PEOT/PBT) block copolymer
(PolyVation B.V.; the weight ratio of PEOT/PBT = 55/45, and the molecular
weight (g/mol) of the starting PEG segments used in the polymerization
process is 300) was dissolved in a mixture of chloroform (CHCl3, Sigma-Aldrich) and 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP,
Biosolve-Chemicals) to prepare the polymer solutions. Solutions of
26–30% of polymer (w/v) in 70:30 to 80:20 CHCl3/HFIP
(v/v) were formulated. Polycaprolactone (PCL, Mn = 80 000, Sigma-Aldrich) solutions were prepared with
20% (w/v) in 75:25 CHCl3/HFIP.
terephthalate/polybutylene terephthalate (PEOT/PBT) block copolymer
(PolyVation B.V.; the weight ratio of PEOT/PBT = 55/45, and the molecular
weight (g/mol) of the starting PEG segments used in the polymerization
process is 300) was dissolved in a mixture of chloroform (CHCl3, Sigma-Aldrich) and 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP,
Biosolve-Chemicals) to prepare the polymer solutions. Solutions of
26–30% of polymer (w/v) in 70:30 to 80:20 CHCl3/HFIP
(v/v) were formulated. Polycaprolactone (PCL, Mn = 80 000, Sigma-Aldrich) solutions were prepared with
20% (w/v) in 75:25 CHCl3/HFIP.
3
Phage KN4-1 Infection Kinetics
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A total of 5 × 103 pfu of phage KN4‐1 was mixed with 5 × 106 cfu of exponential phase culture of 1461 or 4565 strain individually. The culture was continuously incubated at 37°C. Samples were taken at 0.5, 1, 3 and 22 h. After adding CHCl3 (Sigma‐Aldrich, St Louis, MO) to a final concentration of 2% and shaking for 5 min (treated with CHCl3, both intracellular and free phages were determined), the culture was appropriately diluted (or undiluted) with SM buffer, containing 100 mM NaCl, 8 mM MgSO4·7H2O and 50 mM Tris (pH 7.5) (Sigma‐Aldrich, St Louis, MO). For plating, 100 μl of phage suspension was mixed with 100 μl of exponential phase culture of 4565 strain (we use 4565 but not 1461 because phage numbers can be estimated by calculating the visible plaques on 4565). After incubation at 37°C for 10 min, 4 ml 0.7% top agar was added. The mixture was lightly vortexed, poured on a LB agar and incubated at 37°C for 18 h. Phage titres at each time point were determined by plaque counting.
4
Photochemical Characterization of MBB-1
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Absorption spectra were recorded at room temperature (∼25 °C) with a JASCO V-670 spectrophotometer and all solutions were examined in quartz cells with 2-mm pathlength (HELLMA) with a concentration of 2.2 × 10–3 M. Photochemical reaction of MBB-1 was performed in air-equilibrated CHCl3 (Uvasol Merck-Millipore) or in degassed diethyalmine (Sigma-Aldrich) by using an Ultraviolet lamp (UV-6 L/M Herolab) with λirr=365 (±5) nm and a power density=1.7 mW cm−2.
5
Preparation of Dexamethasone-loaded Microparticles
6
Synthesis and Catalytic Application of Manganese Porphyrin
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All reagents and solvents were obtained from commercial sources and used without further purification unless otherwise stated. SBA-15 (Sigma-Aldrich) and manganese(III) 5,10,15,20-tetra(4-pyridyl)-21H,23H-porphine chloride tetrakis(methochloride) (1 ) (Sigma-Aldrich) were purchased. Iodosylbenzene (PhIO) was prepared by the hydrolysis48 (link) of iodosylbenzenediacetate (Sigma-Aldrich, 98%). Cyclohexene (Merck) was purified with a short-activated silica column, CHCl3 (≥ 99%, Merck), CH3CN (99.8%, Merck), and DI water (18-20 MΩ•cm-1 at 27 °C) were used.
7
Electrospun MgO/PCL Nanofibrous Scaffolds
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The MgO/PCL nanofibrous scaffolds were fabricated by electrospinning technique. The PCL (MW = 80 000, Sigma Aldrich, St. Louis, MO) granules were dissolved in DMF (Merck, Germany, Cat no: 616-001-00-X): CHCl3 (Merck-Germany, Cat no: K45116345347) (2:8) to obtain a 10% w/v solution and homogeneous mixture. Afterward, MgO powder was gradually added with the ratios of 1:0, 1:0.005, and 1:0.01. Each solution was separately mixed using a vortex for 30 min. The polymeric mixture was filled into a 10-mL plastic syringe with a 20-gauge stainless steel needle, and then electrospinning was carried out. The syringe tip was charged to a supplied high-voltage (18 kV), and the steady-state flow rate was set to 1 mL/h. The polymer solution was deposited as solid fibers onto a fixed grounded aluminum sheet at 10 cm from the needle tip. The collected electrospun nanofibers were subsequently vacuum-dried for at least 48 hours to remove any residual solvents.
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The MgO/PCL nanofibrous scaffolds were fabricated by electrospinning technique. The PCL (MW = 80 000, Sigma Aldrich, St. Louis, MO) granules were dissolved in DMF (Merck, Germany, Cat no: 616-001-00-X): CHCl3 (Merck-Germany, Cat no: K45116345347) (2:8) to obtain a 10% w/v solution and homogeneous mixture. Afterward, MgO powder was gradually added with the ratios of 1:0, 1:0.005, and 1:0.01. Each solution was separately mixed using a vortex for 30 min. The polymeric mixture was filled into a 10-mL plastic syringe with a 20-gauge stainless steel needle, and then electrospinning was carried out. The syringe tip was charged to a supplied high-voltage (18 kV), and the steady-state flow rate was set to 1 mL/h. The polymer solution was deposited as solid fibers onto a fixed grounded aluminum sheet at 10 cm from the needle tip. The collected electrospun nanofibers were subsequently vacuum-dried for at least 48 hours to remove any residual solvents.
8
Corrosion Resistance of Graphene-Reinforced Coatings
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Thiophene monomer (Merck), graphite
powder (sigma), FeCl3 (Merck), titanium(IV) isopropoxide
(TTIP) (sigma), CHCl3 (CDH), N-methyl-2-pyrrolidone
(NMP) (Merck), BADGE epoxy resins (Sigma), triethylene tetraamine
(Merck), HCl (analytical grade), NaCl (Merck), ethanol (analytical
grade), and CTAB (CDH) were used as received. The LCS specimens (substrate)
of the dimension 40.0 × 15.0 × 1.3 mm were used for salt
spray tests and morphological examinations (immersion test), whereas
circular specimens with a 1 cm2 exposed surface area were
used for electrochemical and wettability tests. The chemical composition
(wt %) of LCS is given inTable 6 . Before coating, the steel specimens were ground with
different grit SiC sheets and then washed with acetone, followed by
ethanol and double-distilled water, and finally dried in hot air.
powder (sigma), FeCl3 (Merck), titanium(IV) isopropoxide
(TTIP) (sigma), CHCl3 (CDH), N-methyl-2-pyrrolidone
(NMP) (Merck), BADGE epoxy resins (Sigma), triethylene tetraamine
(Merck), HCl (analytical grade), NaCl (Merck), ethanol (analytical
grade), and CTAB (CDH) were used as received. The LCS specimens (substrate)
of the dimension 40.0 × 15.0 × 1.3 mm were used for salt
spray tests and morphological examinations (immersion test), whereas
circular specimens with a 1 cm2 exposed surface area were
used for electrochemical and wettability tests. The chemical composition
(wt %) of LCS is given in
different grit SiC sheets and then washed with acetone, followed by
ethanol and double-distilled water, and finally dried in hot air.
9
Trace Elements Analysis of Surface Water
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Surface water was sampled for trace elements analysis from October 2011 to November 2012 in 11 gauging cross-sections situated along the reservoirs. The cross-sections were situated in Kowalskie and Stare Miasto reservoirs (Fig. 1 ). Due to the specific shape of retention reservoirs, water for analyses was sampled above the reservoir, as well as above and below the central and main dam. Due to the fact that Stare Miasto reservoir is intersected by the A2 motorway, one sampling point was additionally localized below.
The samples with volume of 100 mL were collected in polyethylene bottles (HDPE) produced by Nalgene® using a Toń 2 sampler (Mera Błonie, Gdańsk, Poland) and preserved with 0.5 mL of 60% HNO3 Ultrapur® and CHCl3 (both Merck; Darmstadt, Germany). The water temperature, pH, and electrolytic conductivity were measured directly at the sampling sites using a multi-function Multi 197i device equipped with the electrodes: pH-Electrode SenTix 41 and TetraCon® 325 produced by WTW (Weilheim, Germany). After sampling, the samples were taken to the chemical laboratory in a mobile refrigerator at a temperature of 4 ± 2.5 °C. Adequate precautions were exercised to avoid contamination of water during sampling, transport, and handling.
The samples with volume of 100 mL were collected in polyethylene bottles (HDPE) produced by Nalgene® using a Toń 2 sampler (Mera Błonie, Gdańsk, Poland) and preserved with 0.5 mL of 60% HNO3 Ultrapur® and CHCl3 (both Merck; Darmstadt, Germany). The water temperature, pH, and electrolytic conductivity were measured directly at the sampling sites using a multi-function Multi 197i device equipped with the electrodes: pH-Electrode SenTix 41 and TetraCon® 325 produced by WTW (Weilheim, Germany). After sampling, the samples were taken to the chemical laboratory in a mobile refrigerator at a temperature of 4 ± 2.5 °C. Adequate precautions were exercised to avoid contamination of water during sampling, transport, and handling.
10
Phytochemical and Antioxidant Evaluation
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Folin—Ciocalteu phenol reagent (FCR); gallic acid; 2, 2-diphenyl−1-picrylhydrazyl hydrate (DPPH); quercetin; ascorbic acid; and DMSO were from Sigma. AlCl3, Na2CO3, CH3COOK, HClO4, CHCl3, NaOH, FeCl3, and NaCl were from Merck (Darmstadt, Germany). Ethanol, methanol, petroleum ether, ethyl acetate, nitric acid, and sulphuric acid were from Janssen Chimica (Beerse, Belgium). Nutrient agar, Mueller-Hinton agar, and chloramphenicol were procured from Oxoid, England. All chemicals were of analytical grade.
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