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17 protocols using milli q advantage a10 purification system

1

Synthesis and Purification of Copper(I) Bromide

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Copper(I) bromide (CuBr, Merck, ≥
98%) is purified in glacial acetic acid by continuous stirring until
the suspension solution is pale white. After that, the acetic acid
is removed, followed by multiple washing cycles with ethanol. Next,
the resulting powder is dried in a vacuum oven (room temperature,
overnight). 3-Sulfopropyl methacrylate potassium salt (SPMAK, 98%),
(3-aminopropyl)triethoxysilane (APTES, 98%), α-bromoisobutyryl
bromide (BiBB, 98%), 2,2′-bipyridyl (≥98%), triethylamine
(TEA, ≥ 98%), and ethyl α-bromoisobutyrate (EBiB, ≥
98%) are purchased from Merck and used as received. Methanol (ACS
reagent) and toluene (ACS reagent) are purchased from Biosolve and
used as received. MilliQ water purified from a MilliQ Advantage A10
purification system (Millipore, Billerica, MA, USA) is used.
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2

Quantitative Determination of Mycophenolic Acid and Metabolites

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LC-MS grade methanol was purchased from J.T Baker (Phillipsburg, NJ, USA). Formic acid and ammonium formate were from Sigma–Aldrich (St. Louis, MO,US). Water was prepared in-house using a Millipore Milli-Q Advantage A10 purification system (Darmstadt, Germany). Liquid MPA and MPAG were purchased from Cerilliant corporation (Round Rock, Texas, USA). MPA-d3, MPAG-d3 and AcMPAG-d3 were from Toronto Research Chemicals (Toronto, Canada). The structure of MAP, MPAG and AcMPAG are shown in Fig. 1. The Chromsystems 6 PLUS1 multilevel calibrator mycophenolic acid/glucuronide in plasma/serum calibration kit (Munich, Germany) was used to evaluate accuracy. For immunoassays, the mycophenolic acid Flex reagent cartridge, calibrators, and controls were all purchased from Siemens Healthcare Diagnostics (Frimley, UK).
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3

Synthesis and Characterization of Compounds

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RDV (purity ≥ 98 %), RMP (purity of 93 %), and RTP (purity of 98 %) were synthesized as previously reported [6 (link),18 (link)]. The structure and purity of each compound were identified by NMR and HPLC-UV. RN (purity of 98 %) was purchased from Target Molecule Corp. (Boston, MA, USA). Adenosine triphosphate (ATP, purity of 99 %), uridine triphosphate (UTP, purity of 96 %), 13C,15N-ATP (purity of 98 %), 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB, purity ≥ 98 %), bis (p-nitrophenyl) phosphate (BNPP, purity ≥ 99 %), tris-(2-carboxyethyl)phosphine hydrochloride (TCEP, purity ≥ 98 %), Boc-Lys(Boc)-OH, ammonium hydroxide (NH3∙H2O, 28 % NH3 in H2O), and ammonium acetate (NH4Ac) were obtained from Sigma-Aldrich (St. Louis, MO, USA). PhosSTOP™ EASYpack (PhosSTOP) was purchased from Roche (Basel, Switzerland). The Invitrosol™ LC/MS Protein Solubilizer Kit was purchased from ThermoFisher (MA, USA). Phosphoric acid (H3PO4, purity ≥ 85 %) and silver nitrate (AgNO3) were obtained from Sinopharm Chemical Regent Co., Ltd. (Shanghai, China). Acetonitrile (ACN), formic acid (FA), and methanol (MeOH) were obtained from Merck (NJ, USA). Ultrapure water was obtained using a Millipore Milli-Q® Advantage A10 Purification System (Millipore, Molsheim, France). All other regents were of the highest quality commercially available.
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4

Characterization of Polyphenol Compounds

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Acetonitrile, sodium carbonate, hydrochloric acid, formic acid, ascorbic acid, gallic acid, Folin-Ciocalteu reagent, catechin, malvidine-3-O-glucoside, trans-piceid, trans-resveratrol, sodium phosphate dibasic dodecahydrate, potassium phosphate monobasic, sodium carbonate and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (Saint-Quentin Fallavier, France). Sodium acetate and trifluoroacetic acid were obtained from Carlo Erba Reagents (Peypin, France), and methanol and phloroglucinol from Biosolve Chimie (Dieuze, France). The solvents used were High Performance Liquid Chromatography (HPLC) grade. Deionized water was obtained from a Milli-Q Advantage A10 purification system from Millipore (Fontenay sous Bois, France). Alpha-glucosidase from Saccharomyces cerevisiae, acarbose and para-nitrophenyl alpha-D-glucopyranoside (p-NPG) were purchased from Analytic Lab (Castelnau-le-Lez, France). Trans-astringin and trans-piceatannol were obtained from Carbosynth (Compton, UK) and ChromaDex (Irvine, CA, USA), respectively. Hopeaphenol, isohopeaphenol, Ɛ-viniferin, δ-viniferin and ω-viniferin were isolated from a grapevine raw shoot. The cis-isomers were obtained using Ultraviolet-C (UV-C) irradiation (254 nm) from trans-isomers.
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5

Reagents

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4″-tetrahydrofurfuryl macrozone was synthesized in our laboratory according to the procedure reported in a previously published article [12 (link)]. HPLC-grade acetonitrile was purchased from Fisher Scientific (Loughborough, UK). HPLC-grade ammonium bicarbonate was obtained from Sigma-Aldrich (St. Louis, MO, USA). Deionized water was produced by Millipore Milli-Q Advantage A10 purification system (Molsheim, France). MS-grade acetonitrile was purchased from Carlo Erba (Val de Reuil, France). Deuterated acetonitrile (99.8%-D) was procured from Euriso-Top SAS (Saint-Aubin Cedex, France).
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6

Protein Binding Assay Reagents

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The AGP (from pooled human serum, 99% pure), periodic acid reagent (H5IO6), glycogen (bovine liver), HSA (Cohn fraction V, 99% globulin free, 99% fatty acid free), amitriptyline, carbamazepine, chloramphenicol, chlorpromazine, disopyramide, imipramine, lidocaine, nortriptyline, S-propranolol, and quinidine were from Sigma-Aldrich (St. Louis, MO, USA). The Nucleosil silica (7 μm particle diameter, 100 or 300 Å pore size) was obtained from Macherey Nagel (Düren, Germany). Reagents for the micro bicinchoninic (BCA) protein assay were from Pierce (Rockford, IL, USA). All other chemicals were of the purist grades available. All solutions were prepared using water from a Barnstead NANOpure system (Dubuque, IA, USA) or a Milli-Q Advantage A10 purification system (Millipore, Billerica, MA, USA) and were filtered through 0.20 μm GNWP nylon membranes from Millipore.
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7

Isolation and Analysis of Bioactive Compounds from Traditional Chinese Herbs

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Six common herbs including Lonicera japonica, Scutellaria baicalensis, Forsythia suspensa, Glycyrrhiza uralensis, Cirsium japonicum, and Andrographis paniculata were selected according to the TCM prescriptions [18] , [19] (link), [20] , and purchased from the Tong-Ren-Tang Company (Beijing, China). All the standard compounds were obtained from Shanghai Standard Technology Co., Ltd. (Shanghai, China). Glutathione (GSH) was obtained from Sigma (Shanghai, China). ODS gel AAG12S50 (12 nm, S-50 µm, YMC Co., Ltd., Japan) was used for column chromatography (CC). All solvents used for CC were of at least analytical grade (Shanghai Chemical Reagents Co., Ltd., Shanghai, China). High-performance liquid chromatography (HPLC)-grade acetonitrile (ACN), methanol (MeOH), acetic acid and formic acid were purchased from Merck (NJ, USA). Dimethyl sulfoxide (DMSO) and ammonium acetate (NH4Ac) were purchased from Sigma Chemical Co. (MO, USA). HPLC-grade water was produced by a Millipore Milli-Q® Advantage A10 Purification System (Millipore, Molsheim, France).
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8

Surface-Initiated Atom Transfer Radical Polymerization

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Methyl methacrylate (MMA,
99%) was separated from its polymerization inhibitor content by an
alumina oxide column. N-Isopropylacrylamide (NIPAM,
≥99%) was purified by heating (40 °C) and recrystallization
(0 °C) in toluene. Copper(I) bromide (CuBr) was cleaned
with acetic acid and subsequently washed with ethanol prior to use.
Copper(II) bromide (CuBr2, 99%), (3-aminopropyl)
triethoxysilane (APTES, 99%), α-bromoisobutyryl bromide (BiBB,
98%), N,N,N′,N″,N‴-penta-methyldiethylenetriamine
(PMDETA, 99%), triethylamine (TEA, 99%), tetraethyl orthosilicate
(TEOS, ≥99%), hydrogen chloride (HCl, 60%), and ammonia (NH4OH, 32%) were obtained from Sigma-Aldrich and used without
purification. Milli-Q water was purified from a Milli-Q Advantage
A10 purification system (Millipore).
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9

Synthesis and Characterization of Macrozones

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3a-macrozone and 3b-macrozone were synthesized in our laboratory according to the procedure described in a previously published article [7 (link)]. Acetonitrile (HPLC-grade) was purchased from Fisher Scientific (Loughborough, UK). Ammonium bicarbonate (HPLC-grade) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Deionized water was produced by Millipore Milli-Q Advantage A10 purification system (Molsheim, France). Deuterated acetonitrile (99.8%-D) was procured from Euriso-Top SAS (Saint-Aubin Cedex, France). Acetonitrile (MS-grade) was purchased from Carlo Erba (Val de Reuil, France).
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10

Thermoresponsive Hydrogel Synthesis

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Materials. N-isopropylacrylamide (NIPAM, Aldrich, 97%) was purified by recrystallization twice from toluene/hexane, and dried under vacuum for 48 h at room temperature. 2-Hydroxyethyl methacrylate (HEMA, Aldrich, 97%) was purified by distillation under reduced pressure. N,N-methylenebis(acrylamide) (MBA, 99%), potassium persulfate (KPS), N,N,N′,N′tetramethylethylenediamine (TEMED), sodium alginate, calcium chloride and poly(vinyl alcohol) (PVA, M w = 89 000-98 000, 99+ % hydrolyzed) were purchased from Aldrich. Water was purified with a Milli-Q Advantage A10 purification system (Millipore, Billerica, MA, U.S.A.).
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