The fixed and decalcified samples were embedded in paraffin after which 10 μm sections were obtained. Sections were deparaffinized with xylene and rehydrated. For glycosaminoglycan (GAG) staining, sections were placed in 1% alcian blue 8GX (Sigma-Aldrich) in 3% glacial acetic acid (Merck, Darmstadt, Germany) for 30 min, thoroughly rinsed in distilled water and then counterstained using Mayer's hematoxylin solution (Sigma-Aldrich) for 10 min. Alternatively, for collagen staining, sections were placed in 0.1% picrosirius red stain (Sigma-Aldrich) in saturated aqueous 1.3% picric acid (Sigma-Aldrich) for 1 h, and subsequently rinsed in 0.1 and 0.5% glacial acetic acid. After staining, all sections were again dehydrated and covered with a coverslip using Entellan (Merck).
Glacial acetic acid
Manufactured by Merck Group
Sourced in Germany, United States, United Kingdom, India, Italy, Spain, Singapore, France, Brazil, Canada, Belgium, Switzerland, Japan, China, Australia, Sao Tome and Principe, Chile, Malaysia, Macao, Finland, Ireland, Sweden, Netherlands, Portugal
Glacial acetic acid is a colorless, odorous, and corrosive liquid used as a laboratory reagent. It has a chemical formula of CH3COOH and a concentration of 99.7% or higher. Glacial acetic acid is commonly used in various analytical and research applications, serving as a solvent, catalyst, and pH modifier.
Automatically generated - may contain errors
Lab products found in correlation
Methanol, by Merck Group (180 mentions)
Sodium hydroxide (naoh), by Merck Group (164 mentions)
Hydrochloric acid (hcl), by Merck Group (130 mentions)
Ethanol, by Merck Group (111 mentions)
Sodium acetate, by Merck Group (82 mentions)
Acetonitrile, by Merck Group (79 mentions)
Gallic acid, by Merck Group (73 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (73 mentions)
Chitosan, by Merck Group (71 mentions)
Formic acid, by Merck Group (67 mentions)
Sodium chloride (nacl), by Merck Group (60 mentions)
Chloroform, by Merck Group (59 mentions)
Sodium carbonate, by Merck Group (55 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (50 mentions)
Acetone, by Merck Group (48 mentions)
Sulfuric acid, by Merck Group (44 mentions)
Folin ciocalteu reagent, by Merck Group (44 mentions)
Quercetin, by Merck Group (42 mentions)
Trichloroacetic acid, by Merck Group (39 mentions)
Ammonium acetate, by Merck Group (39 mentions)
1 110 protocols using glacial acetic acid
1
Staining Tissue Samples for GAG and Collagen
2
Spectrophotometric Determination of PBG
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Cells were homogenized in lysis buffer (0.6 M Tris, 0.1M EDTA, pH 8.2). Then 100 μL sample was mixed with 100 μL fresh Modified Ehrlich reagent (dissolve 1.0 g p-dimethylamino benzaldehyde (Sigma) in 30 mL glacial acetic acid (Sigma) and mix with 8 mL 70% perchloric acid (Sigma), then bring to 50 mL with glacial acetic acid) and incubated for 10 min at 37 ℃. Concentration of PBG was determined spectrophotometrically at 37 °C in absorbance at 555 nm.
3
Reserpine-Induced Mouse Model
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Reserpine (Sigma–Aldrich) dissolved in glacial acetic acid (Sigma–Aldrich) was diluted with sterile saline to a final glacial acetic acid concentration of 0.5%. Mice received intraperitoneal administration of Reserpine (10 mg/kg) Reserpine 24 h before the beginning of experiments.
4
Synthesis of CNT-Polymer Composites
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Multi-walled carbon nanotubes (CNTs) were purchased from Sigma-Aldrich (St. Louis, MO, USA) (product number: 791431, lot MKBT4011V) and used as received. Methyl 2-bromopropionate (MBP, Sigma-Aldrich (St. Louis, MO, USA), ≥99%), styrene (Sigma-Aldrich (St. Louis, MO, USA), ≥99%), N,N,N′,N′′,N′′-pentamethyldiethylenetriamine (PMDETA, Sigma-Aldrich(St. Louis, MO, USA), 99%), anisole (Sigma-Aldrich (St. Louis, MO, USA), 99.7%), glacial acetic acid, methanol, dioxane, tetrahydrofuran (THF), dimethylformamide (DMF), ethanol, ethyl acetate and 1,4-diazabicyclo[2.2.2]octane (Sigma-Aldrich, analytical grade) were used without any purification. Tert-butyl acrylate (Sigma-Aldrich (St. Louis, MO, USA), 98%) was purified over a column of basic aluminum oxide and stored under nitrogen before use. Copper(I)bromide (Sigma-Aldrich (St. Louis, MO, USA), 99%) and copper(I)chloride (Sigma-Aldrich (St. Louis, MO, USA), 99%) were stirred in glacial acetic acid for 5 h, filtered, washed with acetic acid, ethanol and ethyl acetate and dried under vacuum before use.
5
HPLC Analysis of Phospholipid-Cholesterol Product
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Phospholipon® 90 H and cholesterol products were obtained from Shanghai Soyoung Biotech. Inc. (Shanghai, China). The VPA reference standard was provided by Octagon Chemical Limited (Yantai, China). Sartorius membrane D0405-100FT dialysis tubing was provided by Sigma-Aldrich (St. Louis, USA). Disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium chloride, potassium chloride, acetonitrile, methanol, glacial acetic acid, and dichloromethane were obtained from Merck (Darmstadt, Germany). Polycarbonate membranes (0.45 μm) and 0.22-μm cellulose acetate membranes were obtained from Whatman. All other reagents were of analytical grade or HPLC grade from Merck (Darmstadt, Germany). Benzoic acid as an internal standard was purchased from Sigma-Aldrich. methanol (high-performance liquid chromatography [HPLC] grade), acetonitrile (HPLC grade), phosphoric acid 85% (analytical grade), sodium dihydrogen phosphate, acetic acid glacial, ammonium acetate, formic acid, ammonium formate, hydrochloride acid, triethylamine, n-hexane, and ethyl acetate were purchased from Merck. Double distilled water was used (Ikapharmindo).
6
Determination of Nitrite and Nitrate
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Solutions of sodium nitrite
and ammonium chloride were used in the concentrations of 8 and 4 mol
L–1, respectively, prepared with ultrapure water
(type I) of a Milli-Q apparatus (Millipore, Burlington, MA, USA).
Glacial acetic acid was used as a catalyst, and ammonium hydroxide
was used as the NOx absorbing solution.
All reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA)
and had an analytical grade.
and ammonium chloride were used in the concentrations of 8 and 4 mol
L–1, respectively, prepared with ultrapure water
(type I) of a Milli-Q apparatus (Millipore, Burlington, MA, USA).
Glacial acetic acid was used as a catalyst, and ammonium hydroxide
was used as the NOx absorbing solution.
All reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA)
and had an analytical grade.
7
Chromate Reduction Reaction Kinetics
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All solutions were prepared with ultrapure Millipore deionized water obtained via a Milli-Q® Advantage A10® Direct water purification system, with a resistivity of 18.2 MΩ cm at 25.0 °C. The following chemicals were used as received without further purification: potassium chromate (Alfa Aesar, 99.0%), sodium dihydrogen citrate (Alfa Aesar, 99.0%), citric acid-disodium salt sesquihydrate (Alfa Aesar, 99.0%), glacial acetic acid (Millipore, ACS grade), sodium acetate anhydrous (Millipore, ACS grade), glycolic acid (Acros, 99.0%), succinic acid (TCI America, ≥99.0%), disodium succinate anhydrous (Acros, 99.0%), dl -malic acid (TCI America, ≥99.0%), propanoic acid (TCI America, ≥99.0%), potassium propionate (TCI America, ≥98.0%), potassium nitrate (Avantor Performance Materials US, 99.0–100.5%), potassium hydroxide (BDH Chemicals, 85.0%), and hydrochloric acid (BDH Chemicals, 36.5–38%). Potassium chloride (BDH Chemicals, 99.0–100.5%) was recrystallized by slow diffusion of ethanol into a supersaturated KCl solution in water and dried under vacuum before use.
Concentrations of Cr(vi ) stated throughout correspond to the value calculated based on the added K2CrO4. As reported previously,7,21 (link) dichromate is likely present at the higher 7.00 mM Cr(vi ) concentrations used.
Concentrations of Cr(
8
SDS-PAGE Protein Analysis Protocol
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All chemicals were of an analytical grade. Water, methanol (MeOH), formic acid, and acetonitrile (LC/MS grade Optima), chloroform, Pierce™ Trypsin Protease MS Grade, Pierce™ DTT (Dithiothreitol), Bolt® MOPS SDS Running Buffer (20×), mini protein gel NuPAGE™ 4 to 12% Bis-Tris, and 4X Bolt™ LDS Sample Buffer were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Dulbecco’s Modified Eagle’s Medium (DMEM), trypsin, glutamine, phosphate-buffered saline (PBS), and Foetal Bovine Serum (FBS) were obtained from Lonza® (Verviers, Belgium). Ethanol (96%) was purchased from Carlo Erba (Peypin, France). Iodoacetamide and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Barcelona, Spain). Tris(hydroxymethyl)aminomethane and glacial acetic acid were obtained from Merck Milipore® (Burlington, MA, USA, EUA). Coomassie Brilliant Blue R-250 was purchased from BIORAD® (Hercules, CA, USA). A 5× SDS-PAGE Sample Loading Buffer and NZYBlue Protein Marker were purchased from Nzytech® (Lumiar, Portugal).
9
Luxol Fast Blue Myelin Staining
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Sections were rehydrated in 1× PBS for 15 min followed by hydration in 95% EtoH for 15 min. Sections were placed in a cuvette and incubated in 0.1% Luxol fast blue solution (0.1 mg of Luxol fast blue [Waldeck #1B-389], 100 mL of 95% EtOH, and 0.5 mL of glacial acetic acid [Millipore]) at 56°C overnight. Excess stain was removed using 95% EtoH followed by rinse in dH2O. Slides were differentiated in 0.05% lithium carbohydrate solution (0.05 mg of lithium carbonate [Karolinska apoteket #941122], 100 mL of dH2O) for 30 s followed by differentiation in 70% EtoH for 30 s and rinsed in dH2O. Sections were mounted using xylene (VWR) and imaged using a light microscope (Leica Reichert Polyvar 2).
10
Quantification of Bacterial Biofilm Formation
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Two hundred microliters of the standardized inoculum were aseptically added to each well of a 96-well polystyrene tissue culture plate (Falcon BD; Becton, Dickinson & Co.). Negative controls were prepared similarly using TSB only. After 24 h of incubation at 37 °C under static conditions, biofilms were washed twice with PBS (pH 7.2) (Merck KGaA) to remove non-adherent cells and then fixed at 60 °C for 1 h. Biofilm biomass was stained for 5 min with 200 µL Hucker-modified crystal violet [45 ] and air-dried (37 °C, 30 min). Finally, crystal violet was extracted by exposure for 15 min to 200 μL of 33% glacial acetic acid (Merck KGaA). Biofilm biomass was measured as OD492 (Sunrise; Tecan, Milan, Italy). Based on the efficiency in biofilm formation, each strain was classified as follows [21 (link)]: non-biofilm-former (NBF) (OD ≤ ODc); weak biofilm-former (WBF) [ODc < OD ≤ (2xODc)]; moderate biofilm-former (MBF) [(2xODc) < OD ≤ (4xODc)]; and high biofilm-former (HBF) (OD > 4xODc). The cut-off value (ODc) for biofilm formation was defined as the mean OD of negative controls + 3x standard deviation.
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