Cells were incubated with various concentration of bovine serum albumin (BSA; A-9576, Sigma-Aldrich), γ-globulin (Wako Pure Chemical Industries, Tokyo, Japan), or the same amount of phosphate buffered saline (PBS) in serum-free DMEM for one hour before exposure to hydrogen peroxide at 37°C. For HEK 293 cells, human serum albumin (HSA; A9731, Sigma-Aldrich) was used instead of BSA. Cells were then exposed to 500 μM hydrogen peroxide (Wako Pure Chemical Industries) for six hours.
Hydrogen peroxide
Manufactured by Fujifilm
Sourced in Japan, United States
Hydrogen peroxide is a chemical compound with the formula H2O2. It is a colorless liquid with a slightly pungent odor. Hydrogen peroxide is commonly used as an oxidizing agent in various industrial and laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Ethanol, by Fujifilm (6 mentions)
Hydrochloric acid (hcl), by Fujifilm (6 mentions)
Dimethyl sulfoxide (dmso), by Fujifilm (6 mentions)
Sulfuric acid, by Fujifilm (4 mentions)
Acetone, by Fujifilm (4 mentions)
2 propanol, by Fujifilm (3 mentions)
2 4 carboxyphenyl 4 4 5 5 tetramethylimidazoline 1 oxyl 3 oxide carboxy ptio, by Dojindo Laboratories (3 mentions)
Tert butyl hydroperoxide, by Merck Group (3 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (3 mentions)
Hematoxylin, by Muto Pure Chemicals (3 mentions)
Trifluoroacetic acid, by Fujifilm (2 mentions)
Cycloheximide (chx), by Abcam (2 mentions)
Ethanol, by Kanto Chemical (2 mentions)
Sodium dihydrogen phosphate, by Fujifilm (2 mentions)
Sodium hydrogen phosphate, by Fujifilm (2 mentions)
Nitric acid, by Fujifilm (2 mentions)
Ammonia water, by Fujifilm (2 mentions)
Sodium hydrogen carbonate, by Fujifilm (2 mentions)
Syto9, by Thermo Fisher Scientific (2 mentions)
Polystyrene ps beads, by Funakoshi (2 mentions)
67 protocols using hydrogen peroxide
1
Antioxidant Effects of Serum Proteins
2
Optimizing Oxidative Stress Assays
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Quercetin, protoporphyrin IX (PpIX), dimethyl sulfoxide (DMSO) was purchased from Sigma-Aldrich, USA. Ethanol, superoxide dismutase (SOD), catalase, ethidium bromide, hypoxanthine, diethylenetriaminepentaacetic Acid (DTPA), trifluoroacetic acid, acetonitrile, iron(II) perchlorate, hydrogen peroxide, fluorescein, phosphoric acid, Dulbecco’s phosphate-buffered saline (D-PBS) were purchased from FUJIFILM Wako Pure Chemical Corporation., Japan. 3’-(p-aminophenyl) fluorescein (APF) was purchased from Sekisui Medical Co. Ltd., Japan. Dihydroethidium DHE was purchased from Invitrogen, CA. Compound libraries (Core library; for pilot screening) were provided by drug discovery initiative (DDI), the University of Tokyo (https://www.ddi.u-tokyo.ac.jp/en/ ). WST-8 was purchased from Dojin Laboratoies, Japan. Xanthine oxidase was purchased from Calbiochem (USA).
3
Derivation and Culture of Genetically Modified Germ Stem Cells
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Green GS cells were derived from C57BL6/Tg14(act-EGFP-OsbY01) transgenic mice on a DBA/2 background (Kanatsu-Shinohara et al. 2003 (link)). We also derived GS cells from 7- to 10-d-old Nox1 KO, B6.129-Hif1atm3Rsjo/J, or Cdkn1a KO mice, as described previously (Kanatsu-Shinohara et al. 2014b (link)). Myc DKO GS cells have been described (Kanatsu-Shinohara et al. 2016 (link)). For hypoxic culture, GS cells were placed into plastic bags and gassed with 1% O2/5% CO2/94% N2. When cells were cultured under 5% or 10% O2, the CO2 level was maintained at 5%, but the N2 level was reduced to 90% or 85%, respectively. Where indicated, GS cells were cultured with 30 µM hydrogen peroxide (Wako), 100 µM MitoTempo, 2 mM LPA (both from Sigma), and 1 mM apocynin (Tokyo Chemical Industry).
For Cdkn1a and KD, p21-shRNA-1, p21-shRNA-2, and empty control vectors were used (Dr. Sally Temple, Albany Medical College, NY) (Fasano et al. 2007 (link)). A mixture of lentiviral particles was used for transfection. For cDNA overexpression, mouse Mycn cDNA was inserted into the CSII-Ef1a-IRES-Venus vector. Virus particles were prepared as described previously (Shinohara and Kanatsu-Shinohara 2020 (link)). The multiplicity of infection was adjusted to 10.0. AxCANCre was used for CRE-mediated deletion of target genes in GS cells (MOI = 2.0).
For Cdkn1a and KD, p21-shRNA-1, p21-shRNA-2, and empty control vectors were used (Dr. Sally Temple, Albany Medical College, NY) (Fasano et al. 2007 (link)). A mixture of lentiviral particles was used for transfection. For cDNA overexpression, mouse Mycn cDNA was inserted into the CSII-Ef1a-IRES-Venus vector. Virus particles were prepared as described previously (Shinohara and Kanatsu-Shinohara 2020 (link)). The multiplicity of infection was adjusted to 10.0. AxCANCre was used for CRE-mediated deletion of target genes in GS cells (MOI = 2.0).
4
hiPSCs Exposure to Toxicants
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hiPSCs were treated with cycloheximide (final concentrations of 1, 10, or 100 µM; Biovision), hydrogen peroxide (1, 10, or 100 µM; Wako), Cadmium Standard Solution (Cd(NO3)2, 1 µM; Wako), or Arsenic Standard Stock Solution (As2O3, 100 nM; Wako), and then harvested at the indicated times after treatments. cycloheximide, cadmium standard solution, and arsenic standard stock solution were diluted in dimethyl sulfoxide. hydrogen peroxide was diluted in diethylpyrocarbonate-treated water.
5
Catalytic valorization of biomass-derived compounds
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d (+)‐glucose (98 %, Kishida), d (−)‐fructose (99 %, Wako), d (+)‐mannose (99 %, Wako), d (+)‐xylose (99 %, Wako), dl ‐glycelaldehyde (>90 %, Sigma‐Aldrich), glycolic acid (97 %, Wako), formic acid (98 %, Wako), hydrogen peroxide (30 %, Wako), and benzoic acid (99.5 %, Wako) were used for the reactions and the analysis. Calcium oxide (CaO, 99.9 %, Wako), hydroxide (Ca(OH)2, 90 %, Kishida), carbonate (CaCO3, 99.95 %, Wako), and phosphate (Ca3(PO4)2, 98 %, Wako) were used as catalysts. Other metal oxides, MgO (99.9 %, Wako), SrO (95 %, Kishida), BaO (90 %, Wako), Sc2O3 (99.9 %, Kojundo), Y2O3 (99.99 %, Wako), La2O3 (99.99 %, Wako), CeO2 (99.9 %, Wako), TiO2 (anatase, 98.5 %, Wako), Nb2O5 (99.9 %, Kojundo), Ta2O5 (99.9 %, Wako), Cr2O3 (Wako), SnO2 (98 %, Wako) and ZnO (99.9 %, Wako) were also used as catalysts. Mg‐Al hydrotalcite (Mg/Al=3) was prepared by a conventional coprecipitation method[39] using Mg(NO3)2 ⋅ 6H2O (99 %, Wako), Al(NO3)3 ⋅ 9H2O (98 %, Wako), Na2CO3 (99.5 %, Kishida) and NaOH (98 %, Kishida).
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6
Hydrogen Peroxide Cytotoxicity Assay
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Hydrogen peroxide (Wako Pure Chemical, Osaka, Japan) was added to the culture medium at various concentration. Cell viability was measured after 24 hours, by the WST-1 assay, as described above.
7
Synthesis and Characterization of Copper(II) Complex
8
Fabrication and Application of Microfluidic Devices
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All chemical reagents used in this work were of analytical grade and water was purified by a Milli-Q® Reference (Nippon Millipore, Tokyo, Japan) system. ENG hard resin used for fabrication of the on-chip injector and a template for microfluidic device was obtained from Kudo 3D Inc. (CA, U.S.A.). Polydimethylsiloxane (PDMS) prepolymer and curing agent were purchased from Dow Corning Toray Co., Ltd. (Tokyo, Japan). Isopropyl alcohol (IPA), hydrochloric acid, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride, ethanol and acetone were obtained from Kanto Chemical Co., Inc. (Tokyo, Japan). Tris(hydroxymethyl)aminomethane (Tris) was obtained from MP Biomedicals, Inc. (CA, U.S.A.). Food dyes (yellow and blue) were purchased from Kyoritsu Foods Co., Ltd. (Tokyo, Japan). Resorufin sodium salt and bovine serum albumin (BSA) were obtained from Sigma-Aldrich Co. LLC (MO, U.S.A.). Amplex Red was obtained from Thermo Fisher Scientific, Inc. (MA, USA). Hydrogen peroxide and horseradish peroxidase (HRP) were purchased from FUJIFILM Wako Pure Chemical Corp. (Tokyo, Japan).
9
Transparent Visualization of Fish Larvae
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Fish larvae were bleached and transparentized according to previous reports51 (link),52 (link), with several modifications. Formalin (10% formalin solution), potassium hydroxide (KOH), hydrogen peroxide, Triton-X, phosphate-buffered saline (PBS), HEPES, and glycerin were purchased from Wako Pure Chemical Industries Ltd. (Tokyo, Japan). All other chemicals used in this study were of analytical grade. O. javanicus larvae (exposure group n = 6; negative control n = 2) were fixed in 10% formalin at 4 °C for 3 days. After removal of formalin, larvae were incubated in pre-bleaching solution (0.3% hydrogen peroxide in PBS) at room temperature overnight. Pre-bleaching solution was replaced with bleaching solution (3% hydrogen peroxide in PBS) and incubated at room temperature overnight. After bleaching, the bleaching solution was replaced with a tissue transparency solution (5% formalin, 5% Triton X-100, 1% KOH in PBS) and incubated at 42 °C for 24 h. In O. latipes, which has a less pigmented peritoneum than O. javanicus, bleaching was omitted, i.e., the tissue transparency solution was directly added after fixation, and incubated at 42 °C for 48 h. Transparency of each sample was confirmed under the microscope.
10
Synthesis of Lanthanide Complexes
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Lutetium(III) nitrate hydrate (99.999%) was purchased from Aldrich Co., Ltd. Terbium(III) chloride hexahydrate (99.95%), n-butyllithium in n-hexane (1.6 mol/L), and chloroform-d (99.8%) were purchased from Kanto Chemical Co., Inc. Tetrahydrofuran, super dehydrated, with a stabilizer (for organic synthesis), and hydrogen peroxide (30%), sodium sulfate, anhydrous were purchased from Wako Pure Chemical Industries, Ltd. 2,2,6,6-Tetramethyl-3,5-heptanedione (>97%), 2,7-dibromophenanthrene (>98.0%) and chlorodiphenylphosphine (>97.0%) were purchased from Tokyo Chemical Industry Co., Ltd.
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