A 1 mM standard BLM solution was prepared by dissolving an appropriate amount of bleomycin sulfate (Merck, Darmstadt, Germany) in a 0.9% NaCl (saline) solution. This BLM solution was further diluted with saline to obtain BLM solution with a concentration of 10 µM. Sodium acetate and acetic acid used to make an acetate buffer (NaAc—HAc) of pH 4.5, which acted as the supporting electrolyte, were purchased from Merck. There were 1 mM stock solutions of Ni(II), Cd(II), Ca(II), V(V), Fe(III), Mg(II), Cu(II), glucose, ascorbic acid, dopamine, adenine, epinephrine, uric acid and testosterone that were prepared from Merck reagents in deionized water or ethanol (testosterone) and stored at 4 °C in the dark until the influence of interferents was examined. Diethylenetriaminepentaacetic acid (DTPA) was purchased from Merck. Ultra-purified water from a Milli-Q system (Millipore, Livingston, Scotland, UK) was used to prepare the solutions.
Diethylenetriaminepentaacetic acid dtpa
Manufactured by Merck Group
Sourced in United States, Belgium, Germany, Italy
Diethylenetriaminepentaacetic acid (DTPA) is a chemical compound used as a chelating agent in various laboratory applications. It is capable of forming stable complexes with metal ions, making it a useful tool in analytical and purification processes.
Automatically generated - may contain errors
Lab products found in correlation
Ascorbic acid, by Merck Group (3 mentions)
Cytochrome c, by Merck Group (3 mentions)
Milli q system, by Merck Group (2 mentions)
Horseradish peroxidase hrp, by Merck Group (2 mentions)
Mito tempo h, by Enzo Life Sciences (2 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (2 mentions)
Amplex red hydrogen peroxide peroxidase kit, by Thermo Fisher Scientific (2 mentions)
2 3 dimethoxy 1 4 naphthoquinone dmnq, by Merck Group (2 mentions)
2 7 dichlorofluorescein, by Merck Group (2 mentions)
N acetyl l cysteine, by Merck Group (2 mentions)
Xylenol orange, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Catalase, by Merck Group (2 mentions)
Egg yolk emulsion, by Merck Group (2 mentions)
Chloroform, by Merck Group (2 mentions)
Ampicillin, by Merck Group (2 mentions)
Ethylenediaminetetraacetic acid, by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Cacl2, by Merck Group (2 mentions)
41 protocols using diethylenetriaminepentaacetic acid dtpa
1
Preparation and Characterization of Bleomycin Solutions
2
Quantification of Polycyclic Aromatic Hydrocarbons
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The commercial suppliers of the PAH reagents used, along with the respective catalog number and purity, are listed in Table S1 of the Supplementary Material . The stock solutions of the PAHs were prepared at concentrations of 100 or 200 mg/L in acetonitrile.
The Cc (from equine heart, cat. no. C2506) was obtained from Sigma-Aldrich, Algés, Portugal, as well as HRP (cat. no. P6782). Diethylenetriaminepentaacetic acid (DTPA) was from Merck, Darmstadt, Germany (cat. no. D6518, purity ≥ 99%) and hexane from Fisher Chemical, Geel, Belgium (H/0355/17, ≥99%); PC was from Tokyo Chemical Industry, Tokyo, Japan (D4250) and tetraoleoyl CL from Avanti, Alabaster, AL, USA (710335P). All other chemicals used were HPLC or analytical-grade reagents.
The Cc (from equine heart, cat. no. C2506) was obtained from Sigma-Aldrich, Algés, Portugal, as well as HRP (cat. no. P6782). Diethylenetriaminepentaacetic acid (DTPA) was from Merck, Darmstadt, Germany (cat. no. D6518, purity ≥ 99%) and hexane from Fisher Chemical, Geel, Belgium (H/0355/17, ≥99%); PC was from Tokyo Chemical Industry, Tokyo, Japan (D4250) and tetraoleoyl CL from Avanti, Alabaster, AL, USA (710335P). All other chemicals used were HPLC or analytical-grade reagents.
3
Colicin Sensitivity Assay for Salmonella
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Colicin sensitivity was assayed as described (Nedialkova et al., 2014 (link)). Briefly, an overnight culture of colicin-producing Salmonella enterica serovar Typhimurium strain SL1344 (Spriewald et al., 2015 (link)) was diluted to OD600nm = 0.8, 25 μL was spotted in the center of LB agar plates containing 0.5 μg/mL mitomycin C (Sigma-Aldrich) and 100 μM diethylenetriamine pentaacetic acid (DTPA; Sigma-Aldrich), and grown overnight for 20 h at 37°C. To 10 mL of top agar (0.75% agar), 10 μL of an overnight culture of an individual S. Reading isolate (tester isolate) was added and gently vortexed. The top agar containing the S. Reading isolate was poured onto the plate previously spotted with strain SL1344, incubated for 20 h at 37°C, and the formation of an inhibition zone (halo) around colicin-producing SL1344 was measured to determine the sensitivity of each S. Reading isolate.
4
Elemental Quantification in Ultrapure Solutions
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Concentrated nitric acid (65%) and hydrogen peroxide (30%) of ultrapure grade were purchased from Baker (Milano, Italy), diethylenetriaminepentaacetic acid (DTPA) (purity > 99%) was from Sigma-Aldrich (Italy). Ultrapure water 18.2 MΩ cm, produced with an EASYpureII (Thermo, Milano, Italy), was used for all standard solutions and sample preparations. Y, Lanthanoids, Rh (Rhodium) and Re (Rhenium) standard solutions (1000 ± 5 g mL−1) were purchased from BDH, Merck and CPI International (Milan, Italy).
5
Pluronic Micelles for Antioxidant Assays
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Pluronic F127 ((PEO)200(PPO)65(PEO)200, Mw 12600), Pluronic P104 ((PEO)54(PPO)63(PEO)54, Mw 5900), Pluronic P85 ((PEO)52(PPO)40(PEO)52, Mw 4600) and Pluronic F88 ((PEO)208(PPO)39(PEO)208, Mw 11400), were supplied by BASF corporation. Hydrogen tetrachloroaurate (III) trihydrate (HAuCl4·3H2O), PEO (Mw 8000), sodium chloride (NaCl), hydrochloric acid (HCl), sodium hydroxide (NaOH), 4-acetamidophenol (AAP), horseradish peroxidase (HRP), and diethylenetriaminepentaacetic acid (DTPA) were purchased from Sigma-Aldrich, St. Louis, MO. 1-Hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH), EPR buffer, diethyldithiocarbamic acid sodium salt (DETC), and deferoxamine (DF) were purchased from Noxygen Science Transfer and Diagnostics, Elzach, Germany All reagents were used as received without farther purification. All aqueous solutions were prepared using bi-distilled water (Millipore, Billerica, MA, USA).
6
Synthesis and Characterization of NMR Probes
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15N-PDT (4-oxo-2,2,6,6-tetramethylpiperidine-d16-15N-1-oxyl) (Figure 8 ) originated from CDN Isotopes (Pointe-Claire, Canada). Mito-TEMPO-H (2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl triphenylphosphonium chloride) (Figure 7 ) was purchased from Enzo Lifescience (Antwerpen, Belgium). Boscalid (2-chloro-N-(4-chlorobiphenyl-2-yl)nicotinamide), Bixafen (N-[2-(3,4-dichlorophenyl)-4-fluorophenyl]-3-(difluoromethyl)-1-methylpyrazole-4-carboxamide), superoxide dismutase 2 conjugated with polyethylene glycol (PEGSOD2), ATP disodium, diethylenetriaminepentaacetic acid (DTPA), dextran from Leuconostoc Mesenteroides (average MW 60000–76000) and dimethyl sulfoxide (DMSO) were from Sigma-Aldrich (Overijse, Belgium). The cell-permeant 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) came from InvitrogenTM (Waltham, MA, USA).
7
Enzymatic Antioxidant Assay Reagents
8
Mitochondrial Oxidative Stress Assay
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14N-CMH (1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine) and metformin were purchased from Enzo Lifescience (Antwerpen, Belgium). 15N-PDT (4-oxo-2,2,6,6-tetramethylpiperidine-d16-15N-1-oxyl) originates from CDN Isotopes. Rotenone, superoxide dismutase (SOD), superoxide dismutase conjugated with polyethylene glycol (PEGSOD), xanthine oxidase (XO), hypoxanthine (HX), diethylenetriaminepentaacetic acid (DTPA), dextran from leuconostoc mesenteroides (average MW 60000–76000), adenosine triphosphate (ADP), pyruvate, succinate, malate and phorbol 12-myristate 13-acetate (PMA) were from Sigma-Aldrich (Overijse, Belgium). Mitochondrial assay buffer (MAS) concentrated 2 times (MAS 2x) was composed of sucrose (70 mM), mannitol (220 mM), KH2PO4 (10 mM), MgCl2 (5 mM), HEPES (2 mM), EGTA (ethyleneglycol bis (α-aminoethylether)-N,N1-tetraacetic acid, 1 mM) and FA-free BSA (fatty acid free Bovine Serum Albumin, 0.2%). All MAS components were also purchased from Sigma-Aldrich.
9
Quantifying Hydrogen Peroxide in EAC and BE Cells
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Hydrogen peroxide levels were measured following C-PAC treatment of EAC and BE cells using the Amplex Red Hydrogen Peroxide/Peroxidase kit according to the manufacturer’s recommendations (ThermoFisher Scientific, Waltham, MA). JHAD1 and OE19 EAC cells were plated at 10E3 and 15E3 cells/well, respectively in 96-well black walled, clear bottom plates (Greiner Bio One, Monroe, NC). CP-C BE cells were plated at 8E3 cells/well in the same plates. Cells adhered for 26–30 h, washed once with PBS and treated for 6 h with vehicle, C-PAC [50–100 μg/mL], the inducer 2,3-Dimethoxy-1,4-naphthoquinone (DMNQ; 20 μM final concentration; Sigma Aldrich, St. Louis, MO) and the inhibitor diethylene triamine pentaacetic acid (DTPA; 100 μM final concentration; Sigma Aldrich, St. Louis, MO). After 6 h, the supernatant was transferred to a new plate and the cells were washed. The cells, the supernatant from the cells and the reaction-no cells in medium were assayed independently for hydrogen peroxide 30 min after addition of the Amplex Red substrate. Fluorescence was measured using the SpectraMax® i3 with excitation/emission wavelengths of 545–590 nm. A minimum of 4 wells were analyzed for each treatment and the data expressed as mean relative fluorescence units ± SEM.
10
Oxidative Stress Pathway Analysis
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NaCl, D-glucose, KH2PO4, NaF, dihydroethidium (DHE, stock 10 mM in DMSO), Tris-base, Dulbecco's Modified Eagle Media (DMEM), fetal calf serum (FCS), penicillin/streptomycin, 1x minimal essential amino acids, and 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF, stock 10 mM in DMSO) were all purchased from Thermo Fisher Scientific (Waltham, MA, USA). Acetylcholine, phenylephrine, leupeptin, aprotinin, sodium orthovanadate, pepstatin, phenylmethylsulfonyl fluoride (PMSF), NaHCO3, ethylene glycol tetraacetic acid (EGTA), A23187 (calcium ionophore, stock 10 mM in DMSO), sodium deoxycholate, NP40, lucigenin, diethylenetriaminepentaacetic acid (DTPA), apocynin, and endothelial cell growth supplement were all purchased from Sigma-Aldrich (St. Louis, MO, USA). KCl, CaCl2, MgSO4, and ethylene-diaminetetraacetic acid (EDTA) were all purchased from EMD Millipore (Billerica, MA, USA). Sodium dodecyl sulfate (SDS) and NADPH were purchased from VWR (Radnor, PA, USA). Alexa-647-labeled Annexin V were purchased from BioLegend (San Diego, CA, USA). Heparin was purchased through Leo Pharma (Thornhill, ON, Canada). A complete list of all antibodies used and their sources is provided in Table 1 .
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