POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) and DOGS-Ni-NTA (1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)-succinyl] (nickel salt))were purchased from Avanti Polar Lipids (Alabaster, AL, USA). Atto488 fluorescent dye NHS ester was purchased from ATTO-TEC. 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES), phenylmethanesulfonyl fluoride (PMSF), benzamidine hydrochloride, EDTA-free protease inhibitor tablets and TCEP (Tris(2-carboxyethyl)phosphine hydrochloride) were purchased from Sigma-Aldrich (St Louis, MO, USA). Leupeptin and pepstatin were purchased from Roche (Indianapolis, IN, USA). MOPS (3-(N-morpholino)propanesulfonic acid), β-mercaptoethanol, Triton X-100, NaCl and PBS tablets were purchased from Fisher Scientific.
3 n morpholino propanesulfonic acid mops
Manufactured by Thermo Fisher Scientific
Sourced in United States
3-(N-morpholino)propanesulfonic acid (MOPS) is a chemical compound commonly used as a buffering agent in biological applications. It serves to maintain a stable pH environment, typically in the range of 6.5 to 7.9, in various experimental and analytical procedures.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Crystal violet, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Corning (2 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Tris 2 carboxyethyl phosphine hydrochloride tcep, by Merck Group (1 mentions)
Atto488 fluorescent dye nhs ester, by ATTO-TEC (1 mentions)
4 2 hydroxyethyl 1 piperazineethanesulphonic acid hepes, by Merck Group (1 mentions)
Edta free protease inhibitor tablet, by Merck Group (1 mentions)
Pbs tablet, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Dogs nta ni, by Avanti Polar Lipids (1 mentions)
Pepstatin, by Roche (1 mentions)
Leupeptin, by Roche (1 mentions)
Osmium tetroxide, by Electron Microscopy Sciences (1 mentions)
Tris base, by Thermo Fisher Scientific (1 mentions)
Branched polyethylenimine, by Merck Group (1 mentions)
Sodiumhyaluronate ha, by Thermo Fisher Scientific (1 mentions)
2 3 bis 2 methoxy 4 nitro 5 sulfophenyl 2h tetrazolium 5 carboxanilide xtt, by Thermo Fisher Scientific (1 mentions)
Intramedic, by BD (1 mentions)
7 protocols using 3 n morpholino propanesulfonic acid mops
1
Membrane Protein Purification Protocol
2
Doxycycline-Iron Complex Characterization
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Ligation of doxycycline at 0.1 mM with iron (Iron ICP standard in 2% HNO3, Fisher Scientific) was carried out in ultra-pure water and diluted in MOPS (3-(N-morpholino)propane sulfonic acid (Fisher Scientific) at 0.2 M, pH 7.0. To investigate the stoichiometry of iron binding with doxycycline, 10 different molar fractions of the latter, ranging from 0 to 200 µM, were incubated with different ratios of iron, and full-range UV and visible spectra were obtained in MOPS pH 7.0. The maximal absorption difference between ligand (doxycycline) and complex (doxycycline-iron) was observed at 425 nm. Absorbance of each molar fraction at 425 nm was used to produce a Job's plot [27] (link). For susceptibility testing using the doxycycline-iron complex, the ligation solution (1:1 ratio) was kept at room temperature for 1 h, diluted 10 times with PBS pH 7.4, and filter-sterilised before storage at -20 °C. The pH of growth medium supplemented with doxycycline or complex remained neutral. The minimum inhibitory concentration (MIC) assay was then carried out as described above using doxycycline or doxycycline-iron complex.
3
Fabrication and Characterization of Polyelectrolyte Multilayers
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Catheter tubes (polyethylene, PE-100, inner
diameter =0.034 in.) were obtained from Intramedic, Becton Dickinson
& Co. Poly-l -lysine hydrobromide (PLL, MW = 15,000–30,000),
poly-l -glutamic acid Na salt (PGA, MW = 50 000–100 000),
chitosan (CH, medium molecular weight) and branched polyethylenimine
(BPEI, MW = 25 000) were purchased from Sigma-Aldrich. Sodium
hyaluronate (HA, MW = 1 500 000–2 200 000)
was purchased from Acros Organic. 2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide
(XTT) and RPMI 1640 powder containingl -glutamine and phenol
red (without HEPES or Na bicarbonate) were obtained from Invitrogen.
Phosphate-buffered saline (PBS) concentrate was obtained from OmniPur,
and formaldehyde, glutaraldehyde, and menadione were obtained from
Sigma. Tween-20 was obtained from Acros, and osmium tetroxide (4%)
was obtained from Electron Microscopy Sciences. NaCl, Tris-base, and
3-(N-morpholino)propanesulfonic acid (MOPS) were obtained from Fisher
Scientific. All materials were used as received.
diameter =0.034 in.) were obtained from Intramedic, Becton Dickinson
& Co. Poly-
poly-
chitosan (CH, medium molecular weight) and branched polyethylenimine
(BPEI, MW = 25 000) were purchased from Sigma-Aldrich. Sodium
hyaluronate (HA, MW = 1 500 000–2 200 000)
was purchased from Acros Organic. 2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide
(XTT) and RPMI 1640 powder containing
red (without HEPES or Na bicarbonate) were obtained from Invitrogen.
Phosphate-buffered saline (PBS) concentrate was obtained from OmniPur,
and formaldehyde, glutaraldehyde, and menadione were obtained from
Sigma. Tween-20 was obtained from Acros, and osmium tetroxide (4%)
was obtained from Electron Microscopy Sciences. NaCl, Tris-base, and
3-(N-morpholino)propanesulfonic acid (MOPS) were obtained from Fisher
Scientific. All materials were used as received.
4
Synthesis and Evaluation of β-Amino Acid Derivatives
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Fmoc-β-amino acids,
including Fmoc-l -β-homoalanine, Fmoc-l -β-homovaline,
Fmoc-l -β-homoisoleucine, Fmoc-(1S,2S)-2-aminocyclohexane carboxylic acid, Fmoc-l -β-homophenylalanine,
Fmoc-O-tert-butyl-l -β-homotyrosine, Nβ-Fmoc-Nω-Boc-l -β-homolysine,
and Fmoc-Nω-(2,2,5,7,8-pentamethyl-chromane-6-sulfonyl)-l -β-homoarginine were purchased from Chem-Impex International,
Inc. TentaGel S RAM Fmoc, HBTU (O-(benzotriaole-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate), and HOBt·H2O (N-hydroxybenzotrizole monohydrate) were
purchased from Advanced ChemTech. (S)-3-Aminopentanoic
acid was purchased from Sigma-Aldrich for synthesis of Fmoc-(S)-3-aminopentanoic acid (Fmoc-β3-Et-OH).82 RPMI powder (withl -glutamine and phenol
red, without HEPES and sodium bicarbonate) and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) were purchased from
Invitrogen. 3-(N-Morpholino) propanesulfonic acid
(MOPS) was purchased from Fisher Scientific. Phosphate-buffered saline
(PBS) liquid concentrate (10X) was purchased from OmniPur. Menadione
and melittin were purchased from Sigma. Freshly expired human red
blood cells were obtained from the blood bank at University of Wisconsin-Madison
Hospital.
including Fmoc-
Fmoc-
Fmoc-O-tert-butyl-
and Fmoc-Nω-(2,2,5,7,8-pentamethyl-chromane-6-sulfonyl)-
Inc. TentaGel S RAM Fmoc, HBTU (O-(benzotriaole-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate), and HOBt·H2O (N-hydroxybenzotrizole monohydrate) were
purchased from Advanced ChemTech. (S)-3-Aminopentanoic
acid was purchased from Sigma-Aldrich for synthesis of Fmoc-(S)-3-aminopentanoic acid (Fmoc-β3-Et-OH).82 RPMI powder (with
red, without HEPES and sodium bicarbonate) and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) were purchased from
Invitrogen. 3-(N-Morpholino) propanesulfonic acid
(MOPS) was purchased from Fisher Scientific. Phosphate-buffered saline
(PBS) liquid concentrate (10X) was purchased from OmniPur. Menadione
and melittin were purchased from Sigma. Freshly expired human red
blood cells were obtained from the blood bank at University of Wisconsin-Madison
Hospital.
5
Antifungal Screening of Clinical Isolates
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Clinical isolates of all fungal species (Supplementary Table S3 ) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), BEI Resources (Manassas, VA, USA), or the U.S. CDC (Atlanta, GA, USA). Vero cells (strain NR-10385) were acquired from BEI Resources. Amphotericin B (Fisher Scientific, Fair Lawn, NJ, USA), itraconazole (TCI, Ltd., Tokyo, Japan), 5-Fluorocytosine (TCI, Ltd., Tokyo, Japan), and fluconazole (Acros Organics, New Jersey, USA) were acquired from commercial vendors. Both Amphotericin B and fluconazole were dissolved in DMSO to prepare stock 10 mg/mL solutions. Yeast extract peptone dextrose (YPD, Becton, Dickinson, and Company, Sparks, MD, USA), RPMI-1640 (Gibco, Grand, Island, NY, USA), 3-(N-morpholino)propanesulfonic acid (MOPS, Fisher Scientific, Fair Lawn, NJ, USA), phosphate-buffered saline (PBS, Corning, Manassas, VA, USA), minimum essential medium (MEM, Gibco, Grand Island, NY, USA), sodium pyruvate (Sigma-Aldrich, St. Louis, MO, USA), fetal bovine serum (FBS, Corning, Manassas, VA, USA), penicillin-streptomycin (Gibco, Grand Island, NY, USA), crystal violet (Acros Organics, New Jersey, USA), sodium 3′-[1-[(phenylamino)-carbony]-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene-sulfonic acid hydrate (XTT, Sigma-Aldrich, St. Louis, MO, USA), and 96-well plates (CellTreat, Pepperell, MA, USA) were all purchased from commercial vendors.
6
Mitochondrial Function Evaluation Protocol
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Minimum essential medium with Earl's salt and L-glutamine, RPMI 1640 medium with L-glutamine, fetal bovine serum, Pen-Strep (10 000 U/ml penicillin, 10 mg/ml streptomycin), trypsin 10× were supplied by GIBCO, Invitrogen (Auckland, New Zealand). JC-1 was purchased from Molecular Probes, Invitrogen (Oregon, USA). Dimethyl sulfoxide (DMSO), hydrochloric acid 37%, polyethylene glycol 400 (PEG 400) and 2-propanol were purchased from Merck (Hohenbrunn, Germany). Ethylene glycolbis(aminoethylether)-tetra-acetic acid (EGTA), ethylenediaminetetraacetic acid (EDTA), 3-(N-morpholino)propanesulfonic acid (MOPS), potassium chloride, potassium dihydrogen orthophosphate, sodium chloride, sodium hydrogen carbonate, disodium hydrogen orthophosphate anhydrous, sucrose and Tris were purchased Fisher Scientific (Leicestershire, UK). Thiazoyl blue tetrazolium bromide (MTT) was purchased from Amresco (Ohio, USA). Saline (0.9% sodium chloride) was obtained from Duopharma Sdn. Bhd. (Selangor, Malaysia). Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was purchased from Sigma (Steinheim, Germany). Protein assay dye reagent concentrate was obtained from Bio-Rad Laboratories (California, USA). Complex I, Complex II, Complex IV and ATP synthase enzyme activity microplate assay kit were purchased from MitoSciences (Oregon, USA).
7
Antifungal Susceptibility Profiling of C. auris
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C. auris strains were obtained from the BEI Resources (Manassas, VA, USA), and CDC (Atlanta, GA, USA). Media and reagents were provided from the following chemical vendors: crystal violet (Acros Organics, New Jersey, USA), 3-(N-Morpholino) propane sulfonic acid (MOPS) (Fisher Bioreagents, Fairlawn, NJ, USA), phosphate-buffered saline (PBS) (Corning, Manassas, VA, USA), RPMI 1640 (Gibco, Grand, Island, NY, USA), yeast peptone dextrose (YPD) broth (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), and YPD agar (DOT Scientific Inc, Burton, MI, USA). Drugs were obtained commercially as follows: atazanavir and saquinavir (Ambeed, Arlington Heights, IL, USA), chloramphenicol (Sigma-Aldrich, St. Louis, MO, USA), cyclophosphamide (Cayman Chemical, Ann Arbor, MI, USA), posaconazole (Biosynth Carbosynth, San Diego, CA, USA), and ritonavir (TCI America, Portland, OR, USA).
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