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390 protocols using sodium chloride (nacl)

1

Preparing Lysogeny Broth and M9 Medium

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Lysogeny broth (LB) was prepared by dissolving 1% w/v of tryptone (cat# LP0042; Oxoid), 0.5% w/v of yeast extract (cat# LP0021; Oxoid), and 1% w/v of sodium chloride (cat# 27810.364; VWR Chemicals) in Milli‐Q H2O and sterilized by autoclaving. LB agar plates were prepared by supplementing LB medium with agar and 1.5% w/v before autoclaving. M9 medium was prepared according to the Cold Spring Harbor Protocols (M9 minimal medium (standard), 2010) with modifications in the form of 1× M9 salts (stock solution prepared from 5× M9 salts, see below) supplemented with 0.4% w/v Amicase (cat# 1002372245; Sigma; from 10% w/v sterile‐filtered stock), 0.4% w/v d‐glucose (cat# 101176K; VWR Chemicals; from 40% w/v autoclaved stock), 2 mM MgSO4 (cat#. 25.165.292; VWR Chemicals), 1 µg/ml thiamine (cat# T1270; Sigma), and 100 µM CaCl2 (cat# C3306; Sigma). The 5× M9 salt solution was prepared by dissolving 25.6 g Na2HPO4 (cat# S9390; Merck), 6 g KH2PO4 (cat# P0662; Sigma), 1 g NaCl (cat# 277810.364; VWR Chemicals), and 2 g NH4Cl (cat# 5470.1; Roth) in 400 ml Milli‐Q H2O supplemented with 20 µl of a 10 mg/ml FeSO4 solution (cat# F8048; Sigma). After confirming a pH of 7.4, the solution was sterile‐filtered and stored in the dark.
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2

Formulation and Characterization of Microparticles

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Sodium chloride and chloroform were obtained from VWR (Radnor, PA, USA), whereas 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and trifluoroacetic acid were acquired from Roth (Karlsruhe, Germany). Eudragit ® S100 was purchased from Evonik Industries (Darmstadt, Germany), and PEG (12 kDa) was obtained from FLUKA, part of Fischer Scientific (Hampton, NH, USA). Chitocoat ® for chitosan coating of microspheres was obtained from Freund Corporation (Tokyo, Japan). Lyso-PC (purity: 80 %) and phosphatidylcholine (purity: 99.1 %), both from soybean oil, were kindly donated from Lipoid (Steinhausen, Schwitzerland). Chlorophyll Cl, erythrosine and tartrazine were from Caldic (Rotterdam, Netherlands).
chloroform and Sodium chloride were purchased from VWR (Radnor, PA, USA). All other chemicals were bought from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was provided from an Ultra Clear water system produced by Siemens (Munich, Germany).
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3

Optimized Reactive Oxygen Assay

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PBS was prepared
using 8.77 g/L NaCl (VWR Chemicals, AnalaR Normapur), 1.28 g/L Na2HPO4, 1.36 g/L KH2PO4 (EMSURE,
anhydrous for analysis), 370 μL/L 50% NaOH (Emsure, 50%, for
analysis), and ultrapure water (18.2 MΩ cm, MilliPore, Solna,
Sweden. The pH was adjusted to 7.4. The ionic strength was approximatley
0.23 M.
Saline was prepared using 8.77 g/L NaCl (VWR Chemicals,
AnalaR Normapur) and ultrapure water, and the pH was adjusted to 7.4
with 0.01 M NaOH. The ionic strength was approximatley 0.15 M.
DCFH-DA (Sigma-Aldrich, ≥97%) was diluted in dimethyl sulfoxide
(DMSO) (Sigma-Aldrich, ReagentPlus, ≥99.5%). DMEM, without
phenol red ([+] 4.5 g/L d-glucose, [−] l-glutamine,
[−] pyruvate), was purchased as a sterile solution from Gibco,
Thermo Fisher Scientific, Stockholm, Sweden.
Peroxidase from
horseradish (Sigma-Aldrich, type II, essentially
salt-free, lyophilized powder) was diluted in the specific medium
of interest.
NiCl2, MgCl2 (Sigma-Aldrich,
powder and chunks,
≥99% trace metal basis), and CuCl2 salts were used
for the nanoparticle tracking analysis (NTA) investigation (see below).
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4

Preparing Fe-EDTA Contrast Solution

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Contrast solutions were prepared as 8 g Fe-EDTA (Sigma-Aldrich, St. Louis, MO, USA) and 24 g NaCl (VWR Chemicals) to 368 g demineralized water, with Fe-EDTA dissolved before adding NaCl.
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5

Comprehensive Analytical Characterization

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Chemicals and reagents were of analytical grade and acquired from Merck (Darmstadt, Germany) (Kjeldahl tablets, sulfuric acid, absolute ethanol, sodium carbonate decahydrate, sodium hydroxide, n-hexane and anhydrous sodium sulfate) and Sigma-Aldrich (St. Louis, MO, USA) (Folin–Ciocalteu’s reagent, gallic acid, catechin, heptahydrate ferrous sulfate, 2,2-diphenyl-1-picrylhydrazyl radical, 2,2′-azino-bis(3-ethylenbenzothiazoline-6-sulfonic acid, Trolox, ferric chloride, 2,4,6-tripyridyltriazine, sodium nitrite, aluminum chloride, sodium acetate, hydroxytyrosol and Supelco 37 FAME Mix). Boric acid 4% was purchased from Panreac (Barcelona, Spain) and methanol from Honeywell International, Inc. (Morris Plains, NJ, USA). Sodium chloride and chloroform were acquired from VWR Chemicals (Alfragide, Portugal). The tocopherol and tocotrienol standards and tocol were obtained from Calbiochem (La Jolla, CA, USA) and Matreya Inc. (State College, PA, USA), respectively. The solvents (HPLC grade) were acquired from Chem-Lab (Zedelgem, Belgium) or Merck (Darmstadt, Germany). A Milli-Q water purification system (Millipore, Bedford, MA, USA) was used to treat and obtain the ultrapure water.
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6

Spectrophotometric Analysis of Inorganic Compounds

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All
materials were used
in their original form as received from commercial suppliers. Sodium
chloride (99.5–100%), sulfuric acid (37%), nitric acid (90%),
calcium chloride (95%), sodium bicarbonate (98%), and sodium sulfate
(≥98%) were purchased from VWR Chemicals. Sodium phosphate
(96%), m-xylene (98%), potassium phosphate tribasic
(97%), calcium phosphate (>96%), and magnesium phosphate (98%)
were
supplied by Sigma-Aldrich. DMSO (99%) used was purchased from Alfa
Aesar. All the salts used contained a common sodium cation in order
to eliminate any possible deviations induced by the use of different
cationic species. A Shimadzu UV-3600 UV–vis–NIR spectrophotometer
was used for the measurement and recording of UV spectra.
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7

Synthesis and Analysis of Glucose Derivatives

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3-mercaptophenylboronic acid (3-MPBA) (≥95%), 3-hydroxythiophenol (3-HTP) (≥96%), D-(+)-glucose (≥99.5% GC grade), D-(−)-fructose (≥99%), L-glutamine (≥99%), phorbol 12-myristate 13-acetate (PMA) (≥99%) and sodium hypochlorite (5% w/w) are purchased from Sigma-Aldrich. D-(+)-galactose (≥98%), D-(+)-mannose (≥99%), urea (≥99%), sodium bromide (≥99%), sodium nitroprusside (≥98%), hydrogen peroxide (35% w/w), tert-butyl hydroperoxide (TBHP) (70% w/w), calcium sulfate dehydrate (≥98%) and sodium dihydrogen phosphate (≥98%) are purchased from Alfa Aesar. D-(+)-sucrose (≥99%), creatinine (≥99%) is purchased from TCI. Glucose oxidase from aspergillus and sodium chloride (≥99%) are purchased from VWR. Iron (II) chloride tetrahydrate (≥99%) is purchased from JT Baker. Magnesium sulfate (≥98%) is purchased from EMD Millipore. Potassium chloride (≥99%) is purchased from Acros Organics. Ultrapure water is obtained from a Millipore water system. Citrate-capped gold nanoparticles (60 nm, 5.2×10−11 M) are purchased from Nanopartz. Normal human serum is purchased from UTAK Laboratories. Artificial urine is made from 55 mM sodium chloride, 67 mM Potassium chloride, 2.6 mM calcium sulfate, 29.6 mM sodium sulfate, 3.2 mM Magnesium sulfate, 19.8 mM sodium dihydrogen phosphate, 9.8 mM creatinine and 310 mM urea.
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8

Synthetic PapRIV Characterization

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Synthetic PapRIV was purchased from GL Biochem (Shanghai, China). The alanine scan, metabolites and scrambled control were synthesized using solid-phase peptide synthesis (Supplementary method S1). The quality of all peptides was determined using an in-house developed QC method and a purity of 95% or more was found for each sequence. Calcium dichloride dihydrate, magnesium sulphate, potassium chloride, sodium dihydrogen phosphate hydrate, HEPES, sodium lactate and urethane were purchased from Sigma-Aldrich (Diegem, Belgium), while Bovine Serum Albumin (BSA), sodium iodide, sodium dihydrogen phosphate monohydrate were obtained from Merck KGaA (Darmstadt, Germany). Sodium chloride and disodium hydrogen phosphate dihydrate were obtained from VWR (Leuven, Belgium). Calcium dichloride and d-glucose were purchased from Fluka (Diegem, Belgium) and dextran from AppliChem GmbH (Darmstadt, Germany). Water was purified using an Arium 611 Pro VF purification system (Sartorius, Göttingen, Germany) to laboratory-graded water (18.2 MΩ × cm).
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9

Lipid Characterization Protocols

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All lipids were purchased from Avanti Polar Lipids. Urea, TMAO, chloroform, and methanol were obtained from Sigma-Aldrich. Salt solutions used to control the relative humidity (RH) contained potassium chloride, sodium chloride, and KNO3, which were purchased from VWR Chemicals, or K2SO4, which was purchased from Duchefa Biochemie.
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10

Lipid Extraction and Quantification Protocol

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Hexane, ethanol >99.99%, ethyl acetate, all analytical grade, were obtained from VWR International (Darmstadt, Germany). Methanol, chloroform, methyl acetate, diethyl ether, n-Hexane, potassium chloride, sodium chloride, sulfuric acid, acetic acid and water were of analytical grade and were sourced from VWR International (Darmstadt, Germany). Primuline and acetone were of analytical grade and purchased from Sigma Aldrich. Analytical standard such as triolein (TAG), glyceryl dipalmitate (DAG) and stearic acid (FFA), (PC) phosphatidylcholine, (PE) phosphatidylethanolamine, (Lyso) lysophosphatidylcholine, (PI) phosphatitylinositol, mixture of FAMEs (Fatty Acids Methyl Esters), BSA and glucose were purchased from Sigma Aldrich (Saint-Louis, MO, USA).
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