All chemicals in this study had at least analytical grade quality. Deionized water was obtained from a Purelab ultra (ELGA Labwater, Ede, The Netherlands). Preparation of mobile phase was performed with ammonium acetate solution (7.5 M, Sigma-Aldrich, Steinheim Germany) and glacial acetic acid (Honeywell, Seelze, Germany). IdeS (FabRICATOR®), SpeB (FabULOUS®), and Kgp (GingisKHAN®) proteases were purchased from Genovis (Lund, Sweden). A reference standard therapeutic mAb produced in CHO cells (referred to as mAb1) and the FcɤRIIIa affinity column was obtained from Roche Diagnostics (Penzberg, Germany). An EMA-approved cetuximab (Erbitux®) was used in this study. Cetuximab is a chimeric IgG1, produced by SP2/0 murine myeloma cells, which binds to the epidermal growth factor receptor (EGFR).
Glacial acetic acid
Manufactured by Honeywell
Sourced in Germany, United States
Glacial acetic acid is a concentrated form of acetic acid. It is a clear, colorless liquid with a pungent odor. Glacial acetic acid is commonly used in various laboratory applications as a solvent, reagent, and in the synthesis of other chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Lach-Ner (2 mentions)
Quercetin, by Thermo Fisher Scientific (2 mentions)
Aluminum chloride, by Thermo Fisher Scientific (2 mentions)
Hydrochloric acid (hcl), by Lach-Ner (2 mentions)
Sulfuric acid, by Lach-Ner (2 mentions)
Potassium acetate, by Thermo Fisher Scientific (2 mentions)
Gallic acid standard, by Merck Group (2 mentions)
Sodium acetate trihydrate resistant to potassium permanganate, by Kemika (2 mentions)
Iron 3 chloride hexahydrate, by Kemika (2 mentions)
Trolox standard 6 hydroxy 2 5 7 8 tetramethylchroman 2 carboxylic acid, by Biosynth (2 mentions)
Hplc 99 pure methanol, by Honeywell (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (2 mentions)
Ammonium acetate solution, by Merck Group (1 mentions)
Purelab ultra, by Elga LabWater (1 mentions)
Procainamide hydrochloride, by Merck Group (1 mentions)
Glacial acetic acid, by J&K Scientific (1 mentions)
Sodiumdodecyl sulfate sds, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Pngase f, by Promega (1 mentions)
11 protocols using glacial acetic acid
1
Characterization of Therapeutic Monoclonal Antibodies
2
N-Glycan Labeling and Purification
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N-glycans were released, fluorescently labeled, and cleaned up as described previously [25 (link)], with modification by using procainamide hydrochloride (ProA) as a fluorescent labeling agent for N-glycans. Briefly, isolated IgG was denatured with sodium dodecyl sulfate (SDS) (Invitrogen, USA) and by incubation at 65°C. After enzymatic release with PNGase F (Promega, USA), N-glycans were fluorescently ProA labeled in a 2-step procedure. First, 25 µL of procainamide mixture containing 4.32 mg of procainamide hydrochloride (Sigma-Aldrich) in glacial acetic acid (Honeywell, Charlotte, NC, USA)/dimethyl sulfoxide (Sigma-Aldrich) (30:70) was added per sample, followed by incubation at 65°C for 1 h. Then, 25 µL of reducing agent solution consisting of 4.48 mg of 2-picoline borane (J&K Scientific, Beijing, China) in glacial acetic acid/dimethyl sulfoxide (30:70) was added per sample and incubated at 65°C for an additional 1.5 h. Samples were then cooled for 30 min at RT. Excess reagents were removed from the samples using hydrophilic interaction liquid chromatography–solid phase extraction (HILIC-SPE) as described in [25 (link)]. For the HILIC-SPE step, an AcroPrepadv 1 mL 0.2 μm wwPTFE plate (Pall) was used as the stationary phase. The labeled glycans were eluted with ultrapure water and stored at − 20°C until use.
3
Reagents and Media Sourcing Protocol
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Analytical grade ethyl acetate, methanol, and n-hexane were provided by POCh (Gliwice, Poland), while LC grade acetonitrile, methanol, formic acid and water were acquired from J. T. Baker (Deventer, The Netherlands). Deuterated methanol (CD3OD, 99.90% D) was bought from Eurisotop (Saarbrücken, Germany). 5-Fluorouracil (5-FU), α-melanocyte-stimulating hormone (αMSH), kojic acid, Dulbecco’s phosphate buffered saline (DPBS), 3.3 g/L neutral red solution in DPBS, Dulbecco′s Modified Eagle′s Medium (DMEM), DMEM:F12, Roswell Park Memorial Institute 1640 (RPMI-1640) medium, Ham’s F12 medium, and Eagle′s Minimum Essential Medium (EMEM) were purchased from Sigma-Aldrich (Darmstadt, Germany). Ethanol (>98%) and glacial acetic acid were purchased from Honeywell (Charlotte, NC, USA). Fetal bovine serum (FBS) was obtained from Pan-Biotech (Aidenbach, Germany).
4
Dissolution and Bioanalytical Assay Protocol
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For dissolution testing, analytical reagents were purified water (Milli-Q Elix Essential 5, Merck Chimie SAS, Fontenay-Sous-Bois, France) and hydrochloric acid R (37%) grade for analysis (J.T. Baker). For the bioanalytical assays, TMZ and the internal standard temozolomide-d3 (TMZ-d3) were purchased from TRC (North York, ON, Canada). The reagent-grade acetonitrile, isopropanol and methanol of gradient grade purity were purchased from Carlo Erba, trichloroacetic acid (TCA), ammonium acetate 7.5 M solution and formic acid were purchased from Sigma (St Louis, MO, USA), glacial acetic acid was from Honeywell (Charlotte, NC, USA) and dimethyl sulfoxide (DMSO) was from ACROS. Acetic acid used to prepare acetic acid 10% solution was purchased from Thermo Fisher Scientific (Waltham, MA, USA), and purified water (Milli-Q direct 8, Merck Chimie SAS, Fontenay-Sous-Bois, France) was used. Human plasma from healthy donors was provided by BioIVT (UK). Microtubes in polypropylene were purchased from Sarstedt (Nümbrecht, Germany) and Nunc™ 96 DeepWell™ Polystyrene Plates (DWP) were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
5
Standardized Reagent Preparation Protocol
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Glycerol standard (≥99%) and starch soluble (for analysis) were purchased from Merck (Darmstadt, Germany). Glacial acetic acid (≥99.8%), potassium iodide (≥99.5%), sodium chloride (≥99.5%), sodium hydroxide (≥98%) and buffer solution (pH 4, 7 and 10) were purchased from Honeywell (Charlotte, NC, USA). Sodium acetate trihydrate (99%) was purchased from PENTA chemicals (Prague, Czech Republic) and conductivity standard (1413 μS/cm, 20 °C) was purchased from LLG International (Meckenheim, Germany). Iodine (99.5+) and zinc acetate dihydrate (for analysis) were from Fisher Chemical (Waltham, MA, USA). Potassium hexacyanoferrate (II) trihydrate (98+%) was purchased from Alfa Aesar (Ward Hill, MA, USA). Sodium bisulfite was purchased from Acros Organics (Geel, Antwerp, Belgium).
6
Phytochemical Analysis of Plant Extracts
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HPLC 99% pure methanol obtained from Honeywell (Paris, France) was used as an extraction solvent, while the Folin–Ciocalteu reagent obtained from Fisher Scientific UK (Loughborough, UK) was used for spectrophotometric determination of total phenols. Hydrochloric acid (37%, w/w), sulfuric acid (96%, p.a.), sodium carbonate, anhydrous (99.5–100.5%), and formic acid (98%, p.a.) were obtained from Lach-ner (Neratovice, Czech Republic). Ethanol (96% pure) was obtained from Gram-mol (Zagreb, Croatia). Quercetin (95%), potassium acetate (99%), and aluminum chloride (98.5%) were purchased from Acros Organics (Guangzhou, China). Gallic acid standard (97.5–102.5%), DPPH (2,2-diphenyl-1-picrylhydrazyl radical), and TPTZ (2,4,6-tris-2-pyridyl-s-triazine) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Vanillin (99%), potassium chloride (99.0–100.5%), sodium acetate anhydride (99%), and chlorogenic acid (min. 95%) were purchased from Thermo Fisher (Kandel, Germany). Iron (III)-chloride hexahydrate and sodium acetate trihydrate resistant to potassium permanganate were obtained from Kemika (Zagreb, Croatia). Glacial acetic acid (≥99.8%) was purchased from Honeywell Fluka TM (Seelze, Germany) and trolox standard (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) used for the DPPH and FRAP method was purchased from Biosynth (Bratislava, Slovakia).
7
Comprehensive Analysis of Phytochemicals
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HPLC 99% pure methanol was obtained from Honeywell (Paris, France). Folin-Ciocalteau (FC) reagent was purchased from Fisher Scientific UK (Loughborough, UK). Sodium carbonate, anhydrous (99.5–100.5%), hydrochloric acid (37%, w/w), sulfuric acid (96%, p.a.), and formic acid (98%, p.a.) were purchased from Lach-Ner (Neratovice, Czech Republic). Chlorogenic acid (min. 95%) and vanillin (99%) were purchased from Thermo Fisher (Kandel, Germany). Sodium acetate trihydrate resistant to potassium permanganate and iron (III)-chloride hexahydrate were purchased from Kemika (Zagreb, Croatia). Ethanol (96% pure) was obtained from Gram-mol (Zagreb, Croatia). Potassium acetate (99%), quercetin (95%), and aluminum chloride (98.5%) were obtained from Acros Organics (Guangzhou, China). DPPH (2,2-diphenyl-1-picrylhydrazyl radical), TPTZ (2,4,6-tris-2-pyridyl-s-triazine), and gallic acid standard (97.5–102.5%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Glacial acetic acid (≥99.8%) was obtained from Honeywell FlukaTM (Seelze, Germany), and Trolox standard (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) used for DPPH and FRAP method was obtained from Biosynth (Bratislava, Slovakia).
8
PHB-based Composite Synthesis with Ionic Liquids
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Purified poly(3-hydroxybutyrate) (PHB) powder was provided by the company Bio-On Spa. (Via Santa Margherita Al Colle, 10/3, 40136 Bologna BO, Italy). Ionic liquids 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide 99% (EMIM(TFSI)), 1-buthyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide 99% (BMIM(TFSI)), and choline bis(trifluoromethylsulfonyl)imide 99% (Chol(TFSI)) were purchased from Iolitec GmbH (Salzstraße 184, 74076 Heilbronn, Germany). Tetrabuthylammonium fluoride hydrate 98% (TBAF) was purchased from Sigma Aldrich (Via Gallarate, 154, 20151 Milano MI, Italy) and glacial acetic acid from Riedel-de Haen (Charlotte, North Carolina, NC, US). Gold rods 99.9% were purchased from 8853 Spa (Via Pitagora, 11, 20016 Pero MI, Italy).
9
Equilibrium Solubility of Furosemide in pH 5.2 Buffer
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The equilibrium solubility for FUR was measured in pH 5.2 buffer. A buffer solution of pH 5.2 was prepared according to [25 ] using NaOH (Fisher Scientific UK, Loughborough, UK), NaCl (Fisher Scientific) and glacial acetic acid (Riedel de Haën, Seelze, Germany). The powders (FUR or FUR PM) and 2 mL of buffer were added into the test tubes, which were placed in a shaking water bath (Grant OLS 200, Grant Instruments, Royston, UK) at 37 °C and 100 oscillations/min for three days. The pH was monitored and adjusted to 5.2 with 1 M NaOH or HCl, and powder was added if necessary. The tests were performed in triplicate for each drug and PM. The final solution was filtered through a 45 µm syringe-driven sterile filter (Syringe Filter 30 mm Dia, PES 45 μm Membrane, Sterile, Porvair Sciences), and the drug concentrations were analysed with high-performance liquid chromatography (HPLC) as described below.
10
Characterization of Biological Buffers and Reagents
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