Hydrogen peroxide, ferric sulfate, ferric chloride, catalase, the Hydrogen peroxide assay kit, and the phosphorothioate-modified dinucleotides dGsA and dGA were purchased from Sangon Biotech (Shanghai) Co., Ltd. Trimethoprim, thymine, α-(4-pyridyl-N-oxide)-N-tert-butylnitrone (POBN), diethylenetriaminepentaacetic acid (DTPA), Hanks’ Balanced Salt Solution (HBSS, without phenol red, calcium chloride and magnesium sulfate), threonine, NAD+ and Chelex-100 were purchased from Sigma. The DNeasy Tissue kit was purchased from QIAGEN. The Cycle-Pure Kit, Bacterial DNA Kit, and Gel Extraction Kit were purchased from OMEGA Bio-Tech. Luria-Bertani (LB) broth contained 10 g/L of BactoTryptone, 5 g/L yeast extract, and 10 g/L sodium chloride unless otherwise noted. The modified marine broth medium 2216E contained 5 g/L tryptone, 1 g/L yeast extract, 0.1 g/L FePO4, and 34 g/L sodium chloride. K medium contained A salts, 0.2% glucose, 1 mM MgCl2, 0.5 mM amino acids and 5 mg/L thiamine. The media were supplemented with the following concentrations of antibiotics (unless otherwise noted): ampicillin (Amp) 100 μg/ml and chloramphenicol (Cml) 12.5 μg/ml. Solid medium was supplemented with 1.5% (w/v) agar-A.
Hydrogen peroxide
Manufactured by Sangon
Sourced in China
Hydrogen peroxide is a clear, colorless liquid chemical compound with the formula H2O2. It is a common laboratory reagent used for its oxidizing properties.
Automatically generated - may contain errors
Lab products found in correlation
Stearic acid, by Sangon (2 mentions)
Luciferase assay kit, by Promega (2 mentions)
L arginine, by Sangon (2 mentions)
Ferrous sulfate, by Sangon (2 mentions)
Th f 11 antibody, by Santa Cruz Biotechnology (2 mentions)
Catalase, by Sangon (1 mentions)
Hydrogen peroxide assay kit, by Sangon (1 mentions)
Trimethoprim, by Sangon (1 mentions)
Thymine, by Merck Group (1 mentions)
Threonine, by Merck Group (1 mentions)
Hank s balanced salt solution, by Merck Group (1 mentions)
Chelex 100, by Merck Group (1 mentions)
Dneasy tissue kit, by Qiagen (1 mentions)
Enhanced bicinchoninic acid protein assay kit, by Beyotime (1 mentions)
Lipofectamine 2000, by Merck Group (1 mentions)
L histidine hydrochloride, by Sangon (1 mentions)
Yoyo 1 y3601, by Thermo Fisher Scientific (1 mentions)
L cysteine hydrochloride monohydrate, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride, by Merck Group (1 mentions)
L buthionine sulfoximine, by Merck Group (1 mentions)
11 protocols using hydrogen peroxide
1
Hydrogen Peroxide Assay Protocol
2
Optimizing Transfection Efficiency for Gene Delivery
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Materials used in this study were l -histidine hydrochloride, l -arginine, stearic acid, and hydrogen peroxide (30% w/v; Sangon Biotech, Shanghai, People’s Republic of China); polyethylenimine (PEI, branched, molecular weight 25 kDa), Lipofectamine™ 2000, l -buthionine-sulfoximine, 4′,6-diamidino-2-phenylindole dihydrochloride, l -cysteine hydrochloride monohydrate, ethidium bromide, dithiothreitol (DTT, Sigma-Aldrich, St Louis, MO, USA); a luciferase assay kit (Promega, Madison, WI, USA); pDNA (pGL3 and pEGFP) (Shanghai Innovation Biotechnology Co Ltd, Shanghai, People’s Republic of China); enhanced bicinchoninic acid protein assay kit (Beyotime, Nanjing, People’s Republic of China); Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum, YOYO-1 (Y3601) and penicillin–streptomycin solution 5 kU/mL (Life Technologies, Carlsbad, CA, USA); a Cell Counting Kit-8 (CCK-8, Dojindo Molecular Technologies Inc, Nanjing, People’s Republic of China); and HEK293 cells and HeLa cells (Cell Culture Center of the Shanghai Institutes for Biological Sciences of the Chinese Academy of Sciences, Shanghai, People’s Republic of China). All other reagents were of analytical grade. All animal experiments were performed in accordance with the ethics and regulations of animal experiments of Second Military Medical University (Shanghai, People’s Republic of China).
3
Quantification and Antioxidant Evaluation of Pigments
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Pigment standards including chlorophyll a, violaxanthin, vaucheriaxanthin, and β-carotene were purchased from Sigma-Aldrich Chemical Co. (Shanghai, China; http://www.sigmaaldrich.com ). Silica gel (200–300 mesh) was obtained from Qing Dao Marine Chemical Co. (Qingdao, China). HPLC-grade solvents used for HPLC analysis were purchased from Guangzhou Runhao Biotech Co. (Guangzhou, China), such as methanol, acetonitrile, acetic ether, and dichloromethane. Other analytical solvents (n-hexane, methanol, and acetone) used in extraction and isolation of violaxanthin were purchased from Guangzhou Runhao Biotech Co. (Guangzhou, China). Deionized water was prepared by a Milli-Q water purification system (Millipore Corp., Bedford, MA, USA).
Chemicals used for their antioxidant activity including 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), potassium persulfate, ferrous chloride, potassium ferricyanide, trichloroacetic acid, hydrogen peroxide, ascorbic acid, sodium dihydrogen phosphate, and disodium hydrogen phosphate were obtained from Sangon Biotech Co. (Shanghai, China).
Chemicals used for their antioxidant activity including 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), potassium persulfate, ferrous chloride, potassium ferricyanide, trichloroacetic acid, hydrogen peroxide, ascorbic acid, sodium dihydrogen phosphate, and disodium hydrogen phosphate were obtained from Sangon Biotech Co. (Shanghai, China).
4
Immunohistochemical Analysis of Intestinal Stroma
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Paraffin-embedded tissues were deparaffinized in xylene and rehydrated with descending concentrations of ethanol. Antigen was retrieved in citrate buffer by heating, and nonspecific binding was blocked with 3% hydrogen peroxide (Sangon Biotech Co., Ltd., Shanghai, China) and 5% goat serum (Beyotime Biotechnology, Shanghai, China). The slides were then incubated with specific antibodies at 4°C overnight. After thorough washing with PBS three times, the slides were incubated with an HRP-labelled secondary antibody for 1 h. Diaminobenzidine chromogen was used for the chromogenic reaction. For IF staining, Alexa Fluor 488-conjugated anti-mouse IgG and SABC Cy3-conjugated anti-rabbit IgG were used as secondary antibodies.
The YAP/TAZ and α-SMA positive cells in the intestinal stroma were counted under microscope. The stroma was defined as the population of stromal cells surrounding the identifiable intestinal crypt but did not include identifiable blood/lymphatic vessels or muscular tissue. Ten samples in total were evaluated by immunofluorescence staining. Quantifications were performed with the average values in 3 representative high-power fields per sample. The average number of YAP/TAZ+ stromal cells in every sample was used to analyse the correlation with the number of α-SMA+ cells.
The YAP/TAZ and α-SMA positive cells in the intestinal stroma were counted under microscope. The stroma was defined as the population of stromal cells surrounding the identifiable intestinal crypt but did not include identifiable blood/lymphatic vessels or muscular tissue. Ten samples in total were evaluated by immunofluorescence staining. Quantifications were performed with the average values in 3 representative high-power fields per sample. The average number of YAP/TAZ+ stromal cells in every sample was used to analyse the correlation with the number of α-SMA+ cells.
5
Rhodotorula glutinis SRY Strain Cultivation
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The Rhodotorula glutinis SRY strain was isolated and maintained by the Food Microbiology Team, Agro-Processing Institute, Jilin Academy of Agricultural Sciences. Protein, yeast extract powder, agar powder, sodium chloride, glucose, concentrated hydrochloric acid, acetone, zinc sulfate, manganese sulfate, ferrous sulfate, copper sulfate, hydrogen peroxide and sodium bicarbonate were all purchased from Sangon Biotech (Shanghai) Co., Ltd.; vitamin B1 was purchased from Cisen Pharmaceutical Co., Ltd.; soybean oil was purchased from Yihai Kerry Arawana Holdings Co, Ltd. and TWW is taken from the local market.
6
E. coli Growth and Genetic Manipulation
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E. coli K-12 strains (SI Appendix, Table S1 ) were grown aerobically at 37 °C in LB medium (63 (link)) with shaking at 200 rpm. Colony formation was on LB agar at 37 °C. Bacteriophage P1-mediated transduction (64 ) or CRISPR-based allelic exchange (24 (link)) was used for strain construction. Flow cytometry reagents were purchased from Becton Dickinson. Tryptone, yeast extract, powder for LB broth and agar, and carboxy-H2DCFDA were obtained from Thermo Fisher Scientific. Other reagents, including antimicrobials (ciprofloxacin, oxolinic acid, kanamycin, ampicillin, tetracycline, moxifloxacin, and chloramphenicol), phenol, dimethyl 2-oxoglutarate, cyclic AMP, sodium pyruvate, and DMSO were purchased from Sigma-Aldrich. Gentamicin, amikacin, hydrogen peroxide, chlorhexidine, ethanol, isopropanol, 1-butanol, potassium dichromate, sodium hypochlorite solution (5.2%), hydrogen chloride, and sodium hydroxide were purchased from Sangon Biotech. Meropenem (Shenghuaxi Pharmaceutical) and ceftriaxone (Roche) were from Zhongshan Hospital (Xiamen, China) pharmacy.
7
Investigating Oxidative Stress Modulators
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2FF was provided by SynChem. Inc., Elk Grove Village, IL, USA. Hydrogen peroxide was purchased from Sangon Biotech (Shanghai, China). Catalase (CAT), glutathione (GSH) and malondialdehyde (MDA) test kits were provided by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The mitochondrial membrane potential (MMP) assay kit (JC-1 Kit), lactate dehydrogenase (LDH) kit, MTT cell proliferation and cytotoxicity kit and Mn-SOD activity kit were obtained from Beyotime Biotechnology, China. ROS Fluorescent Probe-Dihydroethidium (DHE) was purchased from Vigorousbio. CO., Ltd. (Beijing, China). Biotinylated aleuria aurantia lectin (AAL, B-1395, 1:3000) was purchased from Vectorlabs, Inc. (Burlingame, CA, USA). Antibodies against Nrf2 (ab62352, 1:1000), NQO1 (ab34173, 1:1000), keap1 (ab139729, 1:2000), NF-κB p65 (ab32536, 1:2000), NF-κB p65 (phospho S536, ab76302, 1:1000) and Nrf2 (phospho S40, ab76026, 1:2000) were from Abcam (Cambridge, UK). Antibodies against glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 60004, 1:5000), HO-1 (10701, 1:2000), cyclooxygenase-2 (COX-2, 66351, 1:1000), IκBα (10268, 1:1000) and Histone (10856, 1:1000) were from Proteintech (Wuhan, China).
8
Luminescent Nanoparticle-based Prostate Cancer Assay
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Enz, aminated GQDs, MAL-PEG-SCM (PEG molecular size: 2000), l -arginine, stearic acid, and hydrogen peroxide (30% w/v) were purchased from Sangon Biotech, Shanghai, China. Dithiothreitol (DTT) was purchased from Sigma-Aldrich (St Louis, MO, USA). A luciferase assay kit was gifted from Promega (Madison, WI, USA). An enhanced bicinchoninic acid protein assay kit was purchased from Beyotime (Nanjing, China). Dulbecco’s Modified Eagle Medium (DMEM), RPMI-1640 medium, foetal bovine serum (FBS), penicillin–streptomycin solution (5 KU/mL) was obtained from Life Technologies (Carlsbad, CA, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies Inc., Nanjing, China. LNCaP and C4-2B cells were obtained from Shanghai Cell Bank, Chinese Academy of Sciences, Shanghai, China. All other reagents were analytical grade. All animals were supplied from Department of Pharmacy of Fudan University and all animal experiments were performed in accordance with the ethics and regulations of animal experiments at Fudan University (Shanghai, China).
9
Osteoblast and Ovarian Cell Responses to Rutin, Iron, and Oxidative Stress
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The hFOB1.19 osteoblast cell line and KGN ovarian granulosa cell line were purchased from the Shanghai Institute of Cell Research, Chinese Academy of Sciences, and cultured at 37 °C with 5% CO2. KGN cells were cultured in DMEM + 10% FBS + 1% double-antibody, while hFOB1.19 cells were cultured in DMEM + 10% FBS + 1% double-antibody. The cultured cells were used for follow-up treatment. The cells were treated with different concentrations of Rutin (0.01 mmol/L, 0.05 mmol/L and 0.1 mmol/L) (Selleck, Houston, TX, USA) for 48 h, followed by 100um ferrous sulfate (Sangon, Shanghai, China) and 600 um hydrogen peroxide (Sangon, China) for one hour. The cells were then collected for follow-up experiments.
10
Immunohistochemistry Protocol for TH and 4-HNE
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Immunohistochemistry was performed as in our previous study64 (link). Briefly, sections were cut into 10-μm slices and antigen retrieval was performed using citrate buffer. Sections were treated with 3% hydrogen peroxide (Sangon Biotech Co., Ltd., Shanghai, China) in PBS for 10 min and were then incubated in 5% BSA for 60 min. Sections were incubated overnight at 4 °C with primary antibodies as follows: TH (F-11) antibody (1:50; Santa Cruz Biotechnology Inc.), and 4-HNE antibody (1:50; R&D Systems.). After washing three times with PBS for 5 min each wash, sections were incubated sequentially in HRP-conjugated goat anti-mouse secondary antibody (ZSGB-BIO, PV6000, Beijing, China) for 1 h at room temperature. Sections were visualized with a 3,3-diaminobenzidine peroxidase substrate kit (ZSGB-BIO, Beijing, China). Integrated optical density was determined using an Image-Pro Plus 6.0 photogram analysis system (IPP 6.0; Media Cybernetics, Bethesda, MD, USA).
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