24 hours after cell plating, the culture media were replaced with a serum-free medium containing 5.5 mM or 25 mM D-glucose. The low glucose culture medium containing 5.5 mM D-glucose was supplemented with 19.5 mM mannitol (Amresco) to adjust the total osmotic pressure to 25 mM. For various purposes, the incubation continued for 6, 24, 48, 72, 96, 120, or 144 hours. As for H/R experiment, cells were incubated in serum-free mediums containing either 5.5 mM or 25 mM D-glucose (the osmotic pressure of which had been balanced as previously stated) for 6 hours before being subjected to hypoxia. At the end of treatment the cells or medium were harvested.
Mannitol
Manufactured by Avantor
Sourced in United States, Belgium
Mannitol is a crystalline polyol compound commonly used as a pharmaceutical excipient in various laboratory applications. It serves as a bulking agent, diluent, and sweetener in formulations. Mannitol exhibits low hygroscopicity and is readily soluble in water, making it a versatile ingredient in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Sucrose, by Avantor (3 mentions)
Acetonitrile, by Merck Group (2 mentions)
Sodium dodecyl sulfate (sds), by Avantor (2 mentions)
Glycerol, by Avantor (2 mentions)
Glycine, by Avantor (2 mentions)
Ammonium persulfate, by Avantor (2 mentions)
Immobilized ph gradient strips, by Bio-Rad (2 mentions)
Bromophenol blue, by Bio-Rad (2 mentions)
Chaps, by Bio-Rad (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (2 mentions)
Pentobarbital sodium, by Solarbio (1 mentions)
Polyvinylidene fluoride membrane, by Bio-Rad (1 mentions)
Urea, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Avantor (1 mentions)
Trifluoroacetic acid tfa, by Merck Group (1 mentions)
Acrylamide, by Bio-Rad (1 mentions)
K3fe cn 6, by Merck Group (1 mentions)
Nycodenz, by Merck Group (1 mentions)
2 de clean up kit, by Bio-Rad (1 mentions)
Butanol, by Merck Group (1 mentions)
11 protocols using mannitol
1
Glucose-dependent Cell Response Dynamics
2
Proteomic Analysis of Mitochondrial Dysfunction
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Acetonitrile, trifluoroacetic acid (TFA), K3Fe(CN)6, HCCA, 5-hydroxydecanote (5-HD), Nycodenz and urea were purchased from Sigma (St. Louis, MO, USA). Bromophenol blue, acrylamide, CHAPS, immobilized pH gradient (IPG) strips, two-dimensional polyacrylamide gel electrophoresis (2DE) Quant kit, 2DE Clean-up Kit, polyvinylidene fluoride membrane and low melting point agarose were purchased from Bio-Rad (Hercules, CA, USA). Glycerol, glycine, ammonium persulfate, N, N′-methylene acrylamide, SDS, Tris, EDTA, sucrose and mannitol were supplied by Amresco (Solon, OH, USA). Secondary antibodies and anti-GRP75, anti-NDUFV1, anti-DLD and anti-cytochrome oxidase IV (COX IV) primary antibodies were obtained from Abcam (Cambridge, UK). TEMED was from Amersham (UK). Butanol was from Merck (Darmstadt, Germany). Trypsin was from Promega (Madison, WI, USA). Pentobarbital sodium and Tween 20 were both from Solarbio (Beijing, China). All other unlisted reagents were of analytical grade or guaranteed reagents.
3
Immunoblot Analysis of Mitochondrial Proteins
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SDS, ammonium persulfate, sucrose, acrylamide, methylene bis-acrylamide, mannitol, glycerol and glycine were purchased from Amresco, LLC. Diazoxide, Nycodenz®, urea, thiourea, EDTA and tetramethylethylenediamine were obtained from Sigma-Aldrich (Merck KGaA). Protein quantification kits, immobilized pH gradient (IPG) strips, dithiothreitol (DTT), BIO-Lyte, CHAPS, agarose, bromophenol blue, β-mercaptoethanol, iodoacetamide and PVDF were acquired from Bio-Rad Laboratories, Inc. Anti-cytochrome c oxidase subunit IV (COX IV; cat. no. ab14744), anti-2-oxoglutarate dehydrogenase (OGDH; cat. no. ab137773), anti-NADH dehydrogenase (ubiquinone) flavoprotein 1 (NDUFV1; cat. no. ab203208) and anti-NADH-ubiquinone oxidoreductase 75 kDa subunit (NDUFS1; cat. no. ab169540) antibodies were purchased from Abcam. All reagents were of analytical grade.
4
Comprehensive LC-MS Analytical Methodology
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LC-MS-grade acetonitrile and ammonium acetate, as well as NH3, were obtained from VWR (Vienna, Austria). Analytical-grade acarbose, arabinose, erythritol, fructose, galactose, glucose, inositol, lactitol, lactose, maltitol, raffinose, rhamnose, ribose, sucrose, xylitol, potassium chloride, ammonium iodide and potassium bromide were purchased from Sigma Aldrich (Schnelldorf, Germany). Erythrose, isomaltulose, lyxose, maltose, maltotriose, mannitol, mannose, sorbitol, sorbose, xylose, sodium nitrate and sodium sulfate were purchased from VWR (Vienna, Austria). LC-MS-grade water (< 0.055 µS cm−1) from an ultrapure water purification system (Sartorius, Göttingen, Germany) was used for both elution and sample preparation. Food samples produced by various companies were obtained from local supermarkets. Sample matrices have been selected over a wide range of beverages and food to investigate the impact on sample preparation and robustness of the analytical method. Food and beverages were stored at the recommended temperature until analysis.
5
Preparation of Plant Growth Regulators
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BAP, GA3 and MeJA were purchased from Sigma-Aldrich (Bornem, Belgium). ABA and ethephon were obtained from Acros Organics (Geel, Belgium). SA, indole acetic acid (IAA) and salt were obtained from Duchefa, whereas mannitol was purchased from VWR (Leuven, Belgium). MG132 was obtained from Enzo Life Sciences (Antwerpen, Belgium). Prior to use, appropriate amounts of MeJA, SA, ABA, IAA and GA3 were dissolved in 100 % (w/v) ethanol, whereas BAP and MG132 were dissolved in 100 % (w/v) dimethyl sulfoxide (DMSO) (VWR). Ethephon, NaCl and mannitol were dissolved in water.
6
Pseudomonas and Staphylococcus Lung Infection
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Pseudomonas aeruginosa (PA01) or Staphylococcus aureus (QO51) was grown in nutrient broth (VWR, Leuven, Belgium) overnight before being refreshed for 3 hr and diluted to OD600 = 0.3 (P. aeruginosa) or OD600 = 5 (S. aureus) in sterile PBS. βENaC-Tg mice (3 weeks old) received a subcutaneous injection of I.6 CatS inhibitor (200 mg/kg) or vehicle control (4% DMSO in peanut oil, Sigma-Aldrich). After 20 min, mice received an intranasal delivery of 20 μl of either P. aeruginosa (OD 0.3), S. aureus (OD 5) or PBS control under isoflurane anaesthetic. Mice were sacrificed 6 hr later, and bronchoalveolar lavage (BAL) fluid collected and processed as described below. Lungs and spleens were collected and homogenised in sterile PBS. Samples were plated on cetrimide (Sigma-Aldrich) (P. aeruginosa) or mannitol (VWR, Leuven, Belgium) (S. aureus) agar for quantification of CFU.
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Pseudomonas aeruginosa (PA01) or Staphylococcus aureus (QO51) was grown in nutrient broth (VWR, Leuven, Belgium) overnight before being refreshed for 3 hr and diluted to OD600 = 0.3 (P. aeruginosa) or OD600 = 5 (S. aureus) in sterile PBS. βENaC-Tg mice (3 weeks old) received a subcutaneous injection of I.6 CatS inhibitor (200 mg/kg) or vehicle control (4% DMSO in peanut oil, Sigma-Aldrich). After 20 min, mice received an intranasal delivery of 20 μl of either P. aeruginosa (OD 0.3), S. aureus (OD 5) or PBS control under isoflurane anaesthetic. Mice were sacrificed 6 hr later, and bronchoalveolar lavage (BAL) fluid collected and processed as described below. Lungs and spleens were collected and homogenised in sterile PBS. Samples were plated on cetrimide (Sigma-Aldrich) (P. aeruginosa) or mannitol (VWR, Leuven, Belgium) (S. aureus) agar for quantification of CFU.
7
Formulation Development of Drug Delivery Systems
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Affinisol HPMC-AS 126
G (>94% purity), Affinisol
HPMC-AS 716 G (>94%), Affinisol HPMC-AS 912 G (>94% purity),
and Methocel
E3 Premium LV Hydroxypropyl Methylcellulose (HPMC E3) were purchased
from Dow Chemicals. Poly(styrene)1.6kDa-block-poly(ethylene glycol)5kDa (PS-PEG) and polycaprolactone3.9 kDa-block-poly(ethylene glycol)5kDa (PCL-PEG) were purchased from Polymer Source Inc. Vitamin E (±-α-tocopherol),
Vitamin E succinate (d -α-tocopherol succinate), pamoic
acid disodium salt (>97% purity), sodium deoxycholate (>97%
purity),
sodium dodecyl sulfate (>99% purity), sodium myristate (>98%
purity),
and sucrose BioXtra (>99.95% purity) were purchased from Sigma-Aldrich.
Tetrahydrofuran (HPLC grade, 99.9%), methanol (HPLC grade, 99.9% purity),
and acetone (HPLC grade, 99.9% purity) were purchased from Fisher
Chemicals. Sodium oleate (>97% purity) was purchased from TCI America.
Phosphate buffered saline 10X (PBS) without calcium or magnesium was
purchased from Lonza.28 Fasted-state simulated
intestinal fluid (FaSSIF), fed-state simulated intestinal fluid (FeSSIF),
and fasted-state simulated gastric fluid (FaSSGF) were purchased from
biorelevant.com.27 Mannitol (>96.0%
purity)
from BDH was purchased from VWR. OZ439 mesylate was supplied by Medicines
for Malaria Venture (MMV).
G (>94% purity), Affinisol
HPMC-AS 716 G (>94%), Affinisol HPMC-AS 912 G (>94% purity),
and Methocel
E3 Premium LV Hydroxypropyl Methylcellulose (HPMC E3) were purchased
from Dow Chemicals. Poly(styrene)1.6kDa-block-poly(ethylene glycol)5kDa (PS-PEG) and polycaprolactone3.9 kDa-block-poly(ethylene glycol)5kDa (PCL-PEG) were purchased from Polymer Source Inc. Vitamin E (±-α-tocopherol),
Vitamin E succinate (
acid disodium salt (>97% purity), sodium deoxycholate (>97%
purity),
sodium dodecyl sulfate (>99% purity), sodium myristate (>98%
purity),
and sucrose BioXtra (>99.95% purity) were purchased from Sigma-Aldrich.
Tetrahydrofuran (HPLC grade, 99.9%), methanol (HPLC grade, 99.9% purity),
and acetone (HPLC grade, 99.9% purity) were purchased from Fisher
Chemicals. Sodium oleate (>97% purity) was purchased from TCI America.
Phosphate buffered saline 10X (PBS) without calcium or magnesium was
purchased from Lonza.28 Fasted-state simulated
intestinal fluid (FaSSIF), fed-state simulated intestinal fluid (FeSSIF),
and fasted-state simulated gastric fluid (FaSSGF) were purchased from
biorelevant.com.27 Mannitol (>96.0%
purity)
from BDH was purchased from VWR. OZ439 mesylate was supplied by Medicines
for Malaria Venture (MMV).
8
Chitosan-Alginate Nanoparticle Synthesis
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Low-molecular-weight chitosan, TPP (MW: 367.864 g/mol) and medium-molecular-weight sodium alginate (viscosity 200–800 cP, 1 wt % in 1% acetic acid, 25 °C) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Trehalose dihydrate and mannitol were purchased from VWR International (Radnor, PA, USA). The water used in this study was of ultrapure quality. All other chemicals and solvents were of analytical grade and were used without further purification.
9
Biochemical Reagent Sourcing for Protein Analysis
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Sucrose and mannitol were purchased from BDH Chemicals and Mallickrodt Chemicals, respectively. Bis-Tris, tricine, and amino-caproic acid were purchased from MB Biochemicals (Irvine, CA). Pre-stained SDS-PAGE markers were purchased from Thermo Scientific (Pittsburgh, PA). Bradford protein assay solution and Coomassie brilliant blue (CBB) R-250 were from Bio-Rad laboratories (Richmond, CA). Silver nitrate, streptozotocin (STZ), sodium citrate, NADH, EDTA, n-dodecyl-β-D-maltoside (DDM), and nitro blue tetrazolium (NBT) chloride tablets were obtained from Sigma (St. Louis, MO, USA). Serva Blue G was purchased from Serva (Heidelberg, Germany). Rabbit anti-HNE polyclonal antibodies (IgG) and goat anti-rabbit IgG conjugated with horseradish peroxidase were purchased from US Biological (Salem, MA) and Invitrogen (San Diego, CA), respectively. Hybond-C membrane and a Western blot detection kit were obtained from GE Healthcare (Piscataway, NJ).
10
Cultivation of Diverse Microorganisms
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