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85 protocols using anhydrous sodium acetate

1

Synthesis of Hematite and Magnetite Nanodiscs

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Synthesis process of nanodiscs follows the protocol described in the previous study11 (link). The synthesis process consists of two steps. First, hematite nanodiscs were synthesized by mixing 10 ml 99.5% ethanol, 0.6 ml ddH2O (or 0.8 ml ddH2O for smaller nanodiscs), 0.8 g of anhydrous sodium acetate (Sigma-Aldrich) and 0.273 g of FeCl3·6H2O (Sigma-Aldrich). After the mixture was homogenized by stirring, the mixture was transferred and sealed into a Teflon-lined steel vessel. The vessel was heated in the oven at 180 °C for 18 h. The hematite nanodiscs were washed with ddH2O twice. Then nanodiscs were washed with ethanol twice. Nanodiscs were dried in a vacuum desiccator. Hematite nanodiscs were further converted into magnetite nanodiscs or used for control experiments. For reduction, 1 mg hematite nanodiscs were mixed with 20 ml of tri-octylamine (Acros Organics) and 1 g of oleic acid (Sigma-Aldrich). The mixture was placed in a three-neck flask connected to a Schlenk line to heated to 370 °C for 25 min in an atmosphere of H2 (5% with 95% Argon, Chiah Lung) and N2 (99.9%, Chiah Lung). During reduction, the red hematite nanodiscs turned to dark gray. After cooled down, the discs were washed with hexane (Alfa Aesar). The magnetite nanodiscs were then dispersed in chloroform (J.T. Baker) and stored in a glass vial at 4 °C.
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2

Extraction and Characterization of Phytochemicals

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Dichloromethane, ethanol, hexane, ethyl acetate, anhydrous sodium acetate, copper (II) sulfate pentahydrate, aluminum potassium sulfate dodecahydrate, calcium carbonate, methanol, aluminum chloride, potassium acetate, Folin-Ciocalteau reagent, sodium carbonate, quercetin, and gallic acid were purchased from Sigma Chemical Co. (Philippines). All the chemicals and reagents were of analytical grade for this study.
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3

Neurotransmitter Metabolite Quantification

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Citric acid monohydrate, anhydrous sodium acetate, sulphatase from Aerobacter aerogenes, pilocarpine and CA standards, such as norepinephrine, DA and isoproterenol (ISO), were purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade methanol was from EMD Millipore (Billerica, MA, USA). Ethylenediamine tetra-acetic acid disodium salt (EDTA.2Na) and sodium 1-octanesulphonate were from Dojindo Laboratories (Kumamoto, Japan). DA-3-sulphate, and DA-4-sulphate was obtained from the NIMH Chemical Synthesis and Drug Supply Program, RTI International.
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4

Acetate Buffer Immobilization Protocol

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A 10-mM solution of immobilization buffer (pH 4.5) was made using anhydrous sodium acetate (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) and acetic acid (Merck Schuchardt OHG, Hohenbrunn, Germany). First, a 0.2-M stock solution was made of both components, then from these stock solutions 1.93 parts of sodium acetate were mixed with 3.07 parts of acetic acid, finally 95 parts of ultrapure demineralized water was added.
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5

Fluorescent Polymer Synthesis Protocol

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7-[4-(Trifluoromethyl)coumarin]methacrylamide), acetonitrile, acrylamide (AAm),
ammonium persulphate (APS), anhydrous sodium acetate (NaOAc), bovine serum albumin
(BSA) ethanol, ethyl acetate, ethylene glycol, ferric chloride hexahydrate
(FeCl3 · 6H2O), fetal calf serum (FCS), fluorescein O-acrylate, glacial acetic acid (AcOH), glutaraldehyde,
myoglobin (Mb) (from equine skeletal muscle), N N,
N
′-methylenebisacrylamide (mBAm), phosphate buffered saline (PBS), sodium
dodecyl sulfate (SDS), tetramethylethyldiamide (TEMED), were all purchased and used
without purification from Sigma-Aldrich, Poole, Dorset, UK.
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6

Optimized Colorimetric Assay Reagents

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NH2OH·HCl (# H9876, ≥99%), AgNO3 (# 209139, ≥99%), oPD (# P9029, ≥98%), DAP (# 661376, 90%), highly stabilized salt-free HRP (# P2088, RZ 2.6–3.4, 200–300 pyrogallol units/mg), NaCl (# S9625, ≥99%), bovine serum albumin (BSA, # A7030, ≥98%), trisodium citrate dihydrate (# S4641, ≥99%), anhydrous sodium acetate (# 1.06268, ≥99%), sodium dihydrogen phosphate (# 71496, ≥99%), sodium phosphate dibasic (# 1.06586, ≥99%), and (3-aminopropyl)-trimethoxysilane (# 281778, 97%) were purchased from Sigma-Aldrich. To minimize the oxidation of oPD, freshly purchased powder was stored at −20 °C instead of recommended +4 °C. Chemically pure NaOH (>99%) for AgNP synthesis and extra-pure grade H2SO4 were purchased from Chimmed, Russia. Chemically pure glacial acetic acid, extra-pure HCl (35–38 weight %), boric acid, and citric acid hydrate were purchased from Component-Reaktiv, Russia. Biotechnology-grade glycine (# Am-O167, >99%) was purchased from Helicon, Russia. Sodium tetraborate decahydrate (# 31457, 99.5%) was purchased from Fluka. All solutions were prepared using deionized water (18.2 MΩ·cm) from the MilliQ UF Plus system (Millipore, Molsheim, France).
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7

Quantification of Osteoclasts in Callus

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One section per sample was stained for tartrate-resistant acid phosphatase (TRAP). Briefly, after deparaffinization and rehydration with distilled water, sections were incubated at 37°C for 25 minutes in a solution of anhydrous sodium acetate (Sigma S-2889), L-(+) tartaric acid (Sigma T-6521), glacial acetic acid, fast red violet LB salt (Sigma F-3381), naphthol AS-MX phosphate (Sigma N-4875), ethylene glycol monoethyl ether (Sigma E-2632), and distilled water. Sections were rinsed in distilled water, counterstained with hematoxylin for 10 sec and then placed in ammonia water for 5 seconds. Quantification was completed using the 10x objective and Osteomeasure software (OsteoMetrics, Inc., Decatur, GA, USA) to contour bone perimeter (B.Pm.) within the anterior callus and identify osteoclasts (Oc.N.), which were defined as multi-nucleated, TRAP-positive cells seated on bone surfaces.
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8

Comprehensive Bioactive Compound Analysis

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HPLC-Grade methanol, hexanes, anhydrous sodium acetate, Folin–Ciocalteu’s phenol reagent, 98% 2,4,6-tripyridyl-s-triazine (TPTZ), 98% (+)-catechin hydrate, butylated hydroxytoluene (BHT), bile extract (from porcine), pepsin (from porcine stomach mucosa) and pancreatin (from porcine pancreas) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Nanopure water was obtained from a Milli-Q System (Millipore Corp., Milford, MA, USA).
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9

Metabolic Profiling of Lactic Acid Bacteria in Skimmed Milk

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L. brevis D6, L. fermentum D12, L. plantarum D13, and E. faecium ZGZA7-10 were inoculated into skimmed milk (2%) (Sigma-Aldrich, Merck, St. Louis, MO, USA) or in skimmed milk with added 2.0, 4.0 and 6.5% (w/v) of NaCl and incubated for 48 h at 37 °C. During cultivation, lactic acid production, pH value, and the bacterial counts were monitored. Proteolytic activity of examined strains after 4 h of incubation in 0.65% (w/v) casein was quantified according to Anson’s method as described by Beganović et al. [38 (link)]. Quantification of lactose, lactate and acetate after overnight growth of particular LAB strains in skimmed milk was performed by liquid chromatography (HPLC-UV/VIS-DAD) while diacetyl was determined by gas chromatography (GC-FID). D-lactose monohydrate (Sigma-Aldrich, Merck, St. Louis, MO, USA), anhydrous sodium acetate (Sigma-Aldrich, Merck, St. Louis, MO, USA), sodium-d,l-lactate (Sigma-Aldrich, Merck, St. Louis, MO, USA), and diacetyl (Sigma-Aldrich, Merck, St. Louis, MO, USA), were used as analytical standards. The quantification limit of the HPLC/UV/VIS (DAD) method is 0.00625 mg/mL for acetate and 0.00625 mg/mL for lactate. The quantification limit for the determination of diacetyl by the GC-FID method is 0.1 mg/mL.
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10

Synthesis of Graphene-Reinforced Nanocomposites

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Ethylene glycol (EG, ≥99%), ethanol (AR, ≥99.5%), iron(iii) chloride (FeCl3·6H2O, AR, 99%), and nickel chloride hexahydrate (NiCl2·6H2O, AR, 98%) were purchased from Aladdin Chemicals, China. Trisodium citrate dihydrate (AR, 99%), anhydrous sodium acetate (AR, ≥99%), melamine (99%), and polyethylene glycol (PEG, average Mn 4000) were purchased from Sigma-Aldrich Chemicals. Ammonia solution (NH3·H2O, 28 wt% in H2O) and tetraethyl orthosilicate (TEOS, reagent grade, 98%) were acquired from Shanghai Macklin Biochemical. Deionized (DI) water, which was used in various processes, was synthesized in our laboratory using a Shenzhen Pure Water No. 1 Water Treatment Technology system. The resistivity of the DI water used in this study was 18.25 mΩ cm−1. The graphene used in the samples was provided by Shanghai Aladdin Biochemical Technology.
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