Hexadecyltrimethylammonium bromide (CTAB, ≥98.0%), hydrogen tetrachloroaurate(III) (HAuCl4·3H2O), sodium borohydride (NaBH4), oleic acid (≥99%), silver nitrate (AgNO3), L-ascorbic acid (99%), a hydrochloric acid solution (HCl, 12 M), thiourea (>99%), molybdic acid (≥85.0% MoO3 basis), 2′,7′-dichlorofluorescein diacetate (H2DCFDA), and ethanol were purchased from Sigma-Aldrich (St. Louis, MO, USA). Phosphate-buffered saline (PBS; 1×, pH 7.4) was purchased from Bioshop (Burlington, ON, Canada). All chemicals were purchased and used without further purification.
2 7 dichlorofluorescein diacetate h2dcf da
Manufactured by Merck Group
Sourced in United States
2',7'-dichlorofluorescein diacetate (H2DCF-DA) is a fluorogenic compound used as an indicator for the detection of reactive oxygen species (ROS) in biological systems. It is a non-fluorescent, cell-permeable compound that undergoes hydrolysis by intracellular esterases, resulting in the formation of 2',7'-dichlorofluorescein (DCF), a highly fluorescent molecule. The intensity of the DCF fluorescence is proportional to the level of ROS present in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
L ascorbic acid, by Merck Group (2 mentions)
α tocopherol, by Merck Group (2 mentions)
N acetyl l cysteine, by Merck Group (2 mentions)
Chloroform, by Merck Group (2 mentions)
Quercetin, by Merck Group (2 mentions)
Methionine, by Merck Group (2 mentions)
2 2 azobis 2 methylpropionamidine dihydrochloride, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (2 mentions)
Ethanol, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Hydrogen tetrachloroaurate, by Merck Group (1 mentions)
Molybdic acid, by Merck Group (1 mentions)
Oleic acid, by Merck Group (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Silver nitrate, by Merck Group (1 mentions)
20 protocols using 2 7 dichlorofluorescein diacetate h2dcf da
1
Synthesis of Gold and Silver Nanoparticles
2
Characterization of TiO2 Nanoparticles
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TiO2 powder was
purchased from Merck. Milling was performed in
a horizontal oscillatory mill (Retsch, PM400) operating at 25 Hz.
Particles were sonicated using a probe sonicator (Vibra-Cell, VCX130,
Sonics, USA) in Milli-Q water medium. TiO2 dispersions
were characterized by the hydrodynamic diameter and zeta potential
by dynamic light scattering (DLS) using a Zetasizer Nano system (Malvern
Instruments, UK). 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium
bromide (MTT) cell culture reagent was purchased from Himedia (India).
2,7-Dichlorofluorescein diacetate (H2DCFDA) was purchased
from Sigma-Aldrich. LipidTOX Deep Red neutral, Alexa Fluor, phalloidin,
Hoechst, and Syto 9 were purchased from Invitrogen (Carlsbad, CA).
Flow cytometry experiments were performed on an Attune acoustic focusing
cytometer (Thermo Scientific, USA).
purchased from Merck. Milling was performed in
a horizontal oscillatory mill (Retsch, PM400) operating at 25 Hz.
Particles were sonicated using a probe sonicator (Vibra-Cell, VCX130,
Sonics, USA) in Milli-Q water medium. TiO2 dispersions
were characterized by the hydrodynamic diameter and zeta potential
by dynamic light scattering (DLS) using a Zetasizer Nano system (Malvern
Instruments, UK). 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium
bromide (MTT) cell culture reagent was purchased from Himedia (India).
2,7-Dichlorofluorescein diacetate (H2DCFDA) was purchased
from Sigma-Aldrich. LipidTOX Deep Red neutral, Alexa Fluor, phalloidin,
Hoechst, and Syto 9 were purchased from Invitrogen (Carlsbad, CA).
Flow cytometry experiments were performed on an Attune acoustic focusing
cytometer (Thermo Scientific, USA).
3
Comprehensive Evaluation of Vegetable and Woody Oils
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Vegetable oils, namely, peanut oil (PNO), soybean oil (SBO), corn oil (CO), flaxseed oil (FO), rapeseed oil (RSO), sunflower seed oil (SUSO), sesame seed oil (SESO), cotton seed oil (CSO), rice bran oil (RBO), pumpkin seed oil (PKSO), hemp seed oil (HSO), perilla seed oil (PLSO), peony seed oil (PNSO), safflower seed oil (SASO), grape seed oil (GSO), and seven woody oil, namely, walnut oil (WO), palm oil (PO), coconut oil (CNO), olive oil (OO), oil-tea seed oil (OSO), yellowhorn seed oil (YSO), sacha inchi oil (SIO) were obtained from local markets.
The Standards for fatty acid methyl esters, α-tocopherol, squalene, campesterol, stigmasterol, and β-sitosterol were from Sigma-Aldrich (St. Louis, MO, USA). The solvents used for the HPLC analysis are chromatography grade from Merck (Darmstadt, Germany). 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), and 2′,7′-dichlorofluorescein diacetate (H2DCF-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Other chemicals and solvents, like Folin-Ciocalteu, sodium carbonate, and potassium persulfate, were analytical grade purchased from Aladdin (Shanghai, China). Superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
The Standards for fatty acid methyl esters, α-tocopherol, squalene, campesterol, stigmasterol, and β-sitosterol were from Sigma-Aldrich (St. Louis, MO, USA). The solvents used for the HPLC analysis are chromatography grade from Merck (Darmstadt, Germany). 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), and 2′,7′-dichlorofluorescein diacetate (H2DCF-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Other chemicals and solvents, like Folin-Ciocalteu, sodium carbonate, and potassium persulfate, were analytical grade purchased from Aladdin (Shanghai, China). Superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
4
Optimized Cellular Assays using Reagents
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Chief purity grade solvents were purchased from Merck, India and Milli-Q water (Milli-Q Academic with 0.22μm Millipak R-40) was used. RPMI-1640, DMEM (Hi-Media, India), fetal bovine serum (Gibco, USA), MTT (Merck, India), NBT (Merck, India), Hoechst (Sigma, USA), Ethidium bromide (EtBr) and Acridine orange (AO) (Merck, India), DMSO (Merck, India), N-acetyl-l-cysteine (NAC) and 2´,7´-dichlorofluorescein diacetate (H2DCFDA) (Sigma, USA), Z-VAD-FMK obtained from Cayman Chemicals (USA), ethanol, chloroform, ethyl acetate, petroleum ether (Merck, India) were used for this study.
5
Fluorescence Microscopy for NO and H2O2 Visualization
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For fluorescence microscopy, NO and H2O2 were visualized using Diaminofluorescein-FM diacetate (DAF-FM-DA) and 2′,7′-dichlorofluorescein diacetate (H2DCF-DA) probes, respectively (Sigma-Aldrich). Briefly, leaf dices were first loaded with 15 μM DAF-FM-DA for 30 min or 50 μM H2DCF-DA for 10 min in Tris/KCl loading buffer (pH 7.2). The process was performed in darkness at 25 °C. Further, leaves were washed three times (5 min each) using Tris/KCl loading buffer (10 mM Tris and 50 mM KCl, pH 7.2), and visualized under a TE2000-U fluorescence microscope (490 nm excitation; 515 nm emission) (Nikon, Tokyo, Japan). Treatments were repeated at least five times. NO and H2O2 contents were also detected using a Griess reagent (Sigma-Aldrich) and an Amplex red hydrogen peroxide/peroxidase assay kit (Invitrogen), respectively [33 (link)].
6
Intracellular ROS Measurement in HUVECs
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The fluorescent probe 2’,7’-Dichlorofluorescein diacetate (H2DCFDA) (Sigma-Aldrich Química, S.L., Madrid, Spain), was used to measure the intracellular production of ROS. Approximately 5×103 HUVECs were grown on clear, flat-bottomed, treated 96-well plates for 21 days under NG, HG or HM conditions. At the end of the experiment, the cells were treated with the indicated drugs, and the reactions were stopped by the staining of cells with 20 mM H2DCFDA for 30 minutes at 37°C. The intensity of H2DCFDA was kinetically measured on a fluorescent microplate reader (Synergy HT, BioTek Instruments, Inc., Winooski, Vermont, USA) in accordance with the manufacturer’s recommendations.
7
Antioxidant Efficacy Evaluation of EGCG-Loaded Lipid Nanocarriers
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Epigallocatechin-3-gallate, phospholipon 90G (high purity soy phosphatidyl choline, SPC), 1,1-diphenyl-2-picrylhydrazyl radical, 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), HA, 2′,7′-dichlorofluorescein diacetate (H2DCFDA) and SC were purchased from Sigma Aldrich, St. Louis, MO. Chloroform, methanol and acetonitrile (HPLC grade) were purchased from Merck Chemicals, Mumbai, India. All other chemicals used were of analytical/reagent grade.
8
Metformin and Cisplatin Synergistic Effects
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Metformin and cisplatin were purchased from Selleck (Huston, USA). 2',7'-Dichlorofluorescein diacetate (H2DCFDA) and N-Acety-L-Cysteine(NAC) was purchased from Sigma (St. Louis, MO, USA). Antibodies against CyclinD1, p21, PARP, caspase3, caspase7, caspase9 and β-Actin were purchased from Cell Signaling Technology (Beverly, MA, USA). The anti-NOX1 antibody was purchased from Abcam (Cambridge, Massachusetts, USA). The Dulbecco’s modified Eagle’s medium (DMEM), Roswell Park Memorial Institute (RPMI1640) and fetal bovine serum were obtained from Gibco (Life Technologies, New York, USA). Crystal violet staining and rhodamin were obtained from Beyotime Institute of Biotechnology (Shanghai, China).
9
Antioxidant Compound Characterization Protocol
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Juglone (5-hydroxy-1,4-naphthoquinone) and 2,7-dichlorofluorescein diacetate (H2DCF-DA) were obtained from Sigma-Aldrich GmbH (Steinheim, Germany). Sodium azide was obtained from AppliChem GmbH (Darmstadt, Germany). EGCG (purity ≥ 95%) was purchased from Sigma-Aldrich (München, Germany). Dimethyl sulfoxide (DMSO), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin-Ciocalteu reagent, and quercetin were obtained from Sigma-Aldrich (MO, USA). Gallic acid was purchased from TCI America (OR, USA). L-Ascorbic acid was obtained from Calbiochem (CA, USA). Other reagents used for extraction were of analytical grade and were purchased from RCI Labscan (Bangkok, Thailand).
10
Oxidative stress assay protocol
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Purpurin (purity ≥ 85.0%), Curcumin (purity ≥ 98.0%), L-glutathione reduced (GSH, purity ≥ 98.0%), L-glutathione oxidized (GSSG, purity ≥ 98.0%), L-buthionine-sulfoximine (BSO, purity ≥ 97.0%), N-acetyl-L-cysteine (NAC, purity ≥ 99.0%), tert-butyl hydroperoxide (TBHP, 70% in water) 2′,7′-dichlorofluorescein (DCF, purity ~90%) and 2′,7′-dichlorofluorescein diacetate (H2DCF-DA, purity ≥ 97.0%) were obtained from Sigma Aldrich (Zwijndrecht, The Netherlands). All buffer salts, including citric acid, tri-sodium citrate dihydrate, di-sodium hydrogen phosphate dodecahydrate, sodium dihydrogen phosphate monohydrate and formic acid, and acetonitrile for Liquid Chromatography/Mass Spectrometry assay, were also obtained from Sigma Aldrich (Zwijndrecht, The Netherlands). Methanol of analytical grade was ordered from Fisher Scientific (Loughborough, United Kingdom). Dimethyl sulfoxide (DMSO) was purchased from Acros Organics (Geel, Belgium). Dulbecco’s Modified Eagle Medium with Ham’s Nutrient Mixture F-12 (1:1) (DMEM/F12), DMEM/F12 without phenol red, trypsin 0.05% EDTA, nonessential amino acids (NEAA) and phosphate-buffered saline pH 7.4 (PBS), geneticin (G418), penicillin/streptomycin, were purchased from Gibco (Carlsbad, CA, USA). Fetal bovine serum (FBS) was purchased from Invitrogen (Breda, The Netherlands).
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