2,2-Diphenyl-2-picrylhydrazyl (DPPH; Aldrich), FeCl2·4H2O (Fluka), trichloroacetic acid (Sigma), phenazine methosulfate (PMS; Sigma), nicotinamide adenine dinucleotide (NADH; Sigma), nitro blue tetrazolium (NBT; Sigma Aldrich), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT; Sigma), butylated hydroxytoluene (BHT; Aldrich), stigmasterol (Sigma), caffeic acid (Sigma), wedelolactone (Sigma), and ethylenediaminetetraacetate (EDTA; Sigma) were purchased from Sigma Chemical Co. (St. Louis, MO). Chlorogenic acid was purchased from Acros Organics (Thermo Fisher Scientific Inc.). Ferrozine, ferric chloride (FeCl3), and potassium ferricyanide (K3Fe (CN)6) were purchased from Showa Co., Ltd. (Tokyo, Japan). Dulbecco's Modified Eagle's Medium (DMEM; Invitrogen), fetal bovine serum (FBS, Gibco), and penicillin-streptomycin were purchased from Gibco BRL (Life technology, Paisley, Scotland).
Nicotinamide adenine dinucleotide nadh
Manufactured by Merck Group
Sourced in United States, Japan
Nicotinamide adenine dinucleotide (NADH) is a coenzyme that plays a vital role in cellular metabolism. It is the reduced form of NAD+, an important electron carrier in many biochemical reactions. NADH is involved in the transfer of electrons during the oxidation-reduction (redox) processes that occur in the mitochondria, where it contributes to the generation of adenosine triphosphate (ATP), the primary energy currency of cells.
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Lab products found in correlation
Trichloroacetic acid, by Merck Group (4 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (4 mentions)
Phenazine methosulfate pms, by Merck Group (3 mentions)
Glutamate, by Merck Group (3 mentions)
Te x band spectrometer, by JEOL (3 mentions)
Succinate, by Merck Group (3 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (3 mentions)
Tris hcl, by Merck Group (3 mentions)
Malate, by Fujifilm (2 mentions)
Nitroblue tetrazolium chloride, by Merck Group (2 mentions)
Ferric chloride, by Merck Group (2 mentions)
Potassium hexacyanoferrate, by Merck Group (2 mentions)
Para methyl styrene, by Merck Group (2 mentions)
Folin ciocalteu s phenol reagent, by Merck Group (2 mentions)
Gallic acid, by Merck Group (2 mentions)
Catechin, by Merck Group (2 mentions)
5 5 dimethyl 1 pyrroline n oxide dmpo, by Dojindo Laboratories (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Nitroblue tetrazolium, by Merck Group (2 mentions)
Phenazine methosulfate, by Merck Group (2 mentions)
15 protocols using nicotinamide adenine dinucleotide nadh
1
Antioxidant and Cytotoxicity Assay Protocol
2
Oxidative Stress Biomarker Analysis
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Trichloroacetic acid (TCA), thiobarbituric acid (TBA), phosphotungstic acid, tetraethoxypropane, nicotinamide adenine dinucleotide (NADH) and phenazine methosulfate (PMS), superoxide dismutase (SOD) standards, ethylenediaminetetraacetic acid (EDTA), nitro blue tetrazolium (NBT), phosphate-buffered saline (PBS), and 1% Triton X-100 were purchased from Sigma–Aldrich (St. Louis, MO, USA). Hydrochloric acid (HCl), acetic acid, sulfuric acid (H2SO4), magnesium chloride (MgC12), sodium hydroxide (NaOH), n-butanol, and hydrogen peroxide (H2O2) were purchased from Fisher Scientific (Chicago, IL, USA). The iron standard solution for AAS was purchased from Fluka (Buchs, Switzerland).
3
Quantifying Intracellular ROS Levels
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Intracellular ROS production levels in HT-treated MCF-7 and MDA-MB-453 cells were measured using electron spin resonance (ESR) in accordance with the protocol described in a previous report [22 (link)]. The cells were seeded on cover slides (49 × 5 × 0.2 mm) and incubated overnight. The cells were incubated at 37 °C or treated at 42 °C for 1 h, and the slides were then immersed in a respiratory solution containing 5 mM succinate (Sigma-Aldrich Co., St. Louis, MO, USA), glutamate (Sigma-Aldrich Co.), malate (Wako Pure Chemical Industries, Ltd.), nicotinamide adenine dinucleotide (NADH) (Sigma-Aldrich Co.), and a 5 μL 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) solution (DOJINDO LABORATORIES). The cell-attached glass cover slide was placed on a tissue glass and inserted into the ESR apparatus. All ESR spectra were obtained using a JEOL-TE Xband spectrometer (JEOL Ltd., Tokyo, Japan) under the following conditions: 7.5 mT sweep width, 1000 gain, 0.1 mT modulation width, and 10 mW incident microwave power. ESR spectra data were analyzed using a Win-Rad Radical Analyzer System (Radical Research Co., Ltd., Tokyo, Japan).
4
Polyphenol Analysis of Natural Compounds
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Folin-Ciocalteu’s phenol reagent, DPPH, ABTS, gallic acid, catechin, nitroblue tetrazolium chloride (NBT), nicotinamide adenine dinucleotide (NADH), Tris-HCl, potassium hexacyanoferrate, trichloroacetic acid, ferric chloride, and para-methyl styrene (PMS) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Polyphenol standards (i.e., chlorogenic acid, p-coumaric acid, rutin, quercetin, and catechin) for HPLC analysis were also purchased from Sigma-Aldrich Co. HPLC-grade water, methanol, acetonitrile, and trifluoroacetic acid (TFA) were purchased from Fisher Scientific Company Llc. (Fair Lawn, NJ, USA). All chemicals used in the described experiments were of an analytical grade.
5
Intracellular ROS Generation Measured by ESR
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Intracellular ROS generation after iron treatment was detected by electron spin resonance (ESR) according to a previous study.(24 (link)) Briefly, cells were seeded on a sterilized glass cover slide (49 × 5 × 0.2 mm) at confluency and incubated overnight. Cells were then exposed to fresh medium containing 500 µM FeSO4 for 1 h and then immersed in the respiratory buffer containing 5 mM succinate (Sigma-Aldrich Japan K.K., Tokyo, Japan), 5 mM malate (Wako), 5 mM glutamate (Sigma-Aldrich Japan K.K.), 5 mM nicotinamide adenine dinucleotide (NADH) (Sigma-Aldrich Japan K.K.) and 5 µl 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) (Dojindo Laboratories, Kumamoto, Japan). The slide was set in a tissue glass, and ESR spectra were obtained using a JEOL-TE X-band spectrometer (JEOL, Ltd., Tokyo, Japan) using the following measurement conditions: 10 mW incident microwave power, 9.4 GHz frequency, and 0.1 mT field modulation amplitude. The experiments were performed three times independently.
6
Antioxidant Activity Evaluation Protocol
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APS was purchased from the Aladdin Industrial Corporation (HPLC purity 95%, Shanghai, China). DPPH, ABTS, phenazine methosulfate (PMS), nicotinamide adenine dinucleotide (NADH), nitroblue tetrazolium (NBT), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), AAPH, BHA, and LPS (from Escherichia coli O55:B5) were purchased from Sigma-Aldrich (St. Louis, MO). Commercial kits used for the determination of enzymes activities of SOD, glutathione peroxidase (GSH-Px), catalase (CAT), and total protein contents were purchased from the Nanjing Jiancheng Bioengineering Institute (Jiangsu, China). All other chemicals used were obtained from the Shanghai chemical agents company, China and were of analytical grade.
7
Mitochondrial Electron Transport Chain Assay
8
ROS Quantification via ESR Spectroscopy
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ROS generation in cells was measured using electron spin resonance (ESR) according to previous study.(13 (link)) Cells were seeded on a glass cover slide (49 × 5 × 0.2 mm) and incubated overnight. Cells were exposed to the medium containing 1 mM IND for 1 h. Cells were immersed in respiratory buffer containing 5 mM succinate (Sigma-Aldrich Japan K.K., Tokyo, Japan), 5 mM malate (Wako Pure Chem. Ind., Ltd., Osaka, Japan), 5 mM glutamate (Sigma-Aldrich Japan K.K.), 5 mM nicotinamide adenine dinucleotide (NADH) (Sigma-Aldrich Japan K.K.), and 10 mM 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) (DOJINDO). The cell-attached glass cover slide was placed on a tissue glass, and the ESR spectra were obtained by inserting the tissue glass into the device. All ESR spectra were obtained using a JEOL-TE X-band spectrometer (JEOL Ltd., Tokyo, Japan) under the following conditions: 20 mW incident microwave power, 9.42 GHz frequency, and 0.1 mT field modulation amplitude.
9
Evaluation of Antioxidant Capacity
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2′,7′-dichlorofluorescein (DCF), DCF-diacetate (DCF-DA), L-KYN, 3-nitropropionic acid (3-NP), FeSO4, xylenol orange, ammonium iron (II) sulfate hexahydrate, 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), bovine serum albumin (BSA), nitroblue tetrazolium (NBT), phenazine methosulfate (PMS), ethylenediaminetetraacetic acid (EDTA), nicotinamide adenine dinucleotide (NADH), diethylenetriaminepentaacetic acid (DTPA), sodium hypochlorite (NaOCl), H2O2 and N,N-dimethyl-4-nitrosoaniline (DMNA) were all obtained from Sigma Chemical Company (St. Louis, MO, USA). All other reagents were reactive grade and obtained from known commercial suppliers. Solutions were prepared using deionized water obtained from a Milli-RQ (Millipore, Burlington, MA, USA) purifier system.
10
Enzymatic Decolorization of Textile Azo Dye
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The textile azo dye Procion Red—H3B (CI No. 31701; molecular formula—C25H15ClN7Na3O10S3; molecular weight—774 g/mol) was gently gifted by Rama International textile Ltd. (Surat, Gujarat, India). For the enzyme assay, nicotinamide adenine dinucleotide (NADH), dichloro phenol indophenol (DCIP), phenanthrene enrichment medium, guaiacol (Sigma-Aldrich, USA), ABTS (2, 20-azino-bis (3-ethyl benzothiazoline- 6-sulfonic acid)) and all other chemicals were purchased from Merck KGaA, Darmstadt, Germany.
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