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Tempol

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Tempol is a nitroxide free radical compound that exhibits antioxidant properties. It scavenges superoxide and other reactive oxygen species, and can modulate cellular redox state. Tempol is commonly used as a research tool in studies investigating oxidative stress and related biological processes.

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Lab products found in correlation

15d pgj2, by Cayman Chemical (2 mentions) Pyrrolidine dithiocarbamate pdtc, by Merck Group (2 mentions) T0070907, by Cayman Chemical (2 mentions) Mk0524, by Cayman Chemical (2 mentions) N 2 mercaptopropionylglycine mpg, by Merck Group (2 mentions) Cay10471, by Cayman Chemical (2 mentions) Gsh ethyl ester, by Merck Group (2 mentions) Dh 15d pgj2, by Cayman Chemical (2 mentions) N acetyl l cysteine, by Merck Group (2 mentions) Pd169316, by Merck Group (2 mentions) Apocynin, by Bio-Techne (1 mentions) Baicalein, by Bio-Techne (1 mentions) α ketobutyrate, by Merck Group (1 mentions) Lactate glo assay, by Promega (1 mentions) Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (1 mentions) Pyruvate assay kit, by Abcam (1 mentions) Gsh detection assay kit, by Abcam (1 mentions) Mitotracker green fm, by Thermo Fisher Scientific (1 mentions) Alpha tocopherol, by Merck Group (1 mentions) Hilyte fluor 488, by AnaSpec (1 mentions)

14 protocols using tempol

1

Amygdalar Modulation of Oxidative Stress and Behavior

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The rat was anesthetized with pentobarbital sodium (50 mg/kg, i.p.) and placed in a stereotaxic apparatus. A craniotomy over the right amygdala was performed and a guide cannula (outer diameter: 0.64 mm; inner diameter: 0.45 mm) was implanted on the dorsal margin of the CeA, using the following coordinates (in mm): 2.5 caudal to bregma; 4.0 lateral to midline; depth, 7.5. The cannula was fixed to the skull with dental acrylic. The behavioral tests were carried out 5 days after the surgery. For drug application, a microinjection tubing probe was inserted through the guide cannula so that the probe protruded by 1 mm. The probe was connected to a PE-10 polyethylene tube filled with dissolved drug solution. The drug solution was pushed into the brain tissue by injecting 1μl air into the polyethylene tube with a microsyringe. Then the polyethylene tube was heat sealed and the probe was left in place for 10 min before the behavioral tests. The following drugs were used: Tempol, baicalein, apocynin (Tocris). Drugs were dissolved in DMSO (30% in deionized water) and diluted in ACSF (containing [in mM] 117 NaCl, 4.7 KCl, 1.2 NaH2PO4, 2.5 CaCl2, 1.2 MgCl2, 25 NaHCO3 and 11 glucose) at 1:100 to their final concentration for injection into the amygdala.
Lu Y.F., Neugebauer V., Chen J, & Li Z. (2016). Distinct contributions of reactive oxygen species in amygdala to bee venom-induced spontaneous pain-related behaviors. Neuroscience letters, 619, 68-72.
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2

Tempol: A Powerful Antioxidant in Neuroscience

Cited 1 time
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4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (tempol, membrane-permeable ROS scavenger, superoxide dismutase mimetic; purchased from Tocris Bioscience) [34 (link);43 (link)] was used in this study. tempol was dissolved in water for stock solutions and diluted in ACSF to the final concentration on the day of the experiment (100-fold that predicted to be needed based on data from our previous studies [29 (link);44 (link)]). Drug concentration in the tissue is at least 100 times lower than in the microdialysis probe as a result of the concentration gradient across the dialysis membrane and diffusion in the tissue. Numbers in the text refer to drug concentrations in the microdialysis fiber.
Ji G., Li Z, & Neugebauer V. (2015). Reactive oxygen species (ROS) mediate visceral pain-related amygdala plasticity and behaviors. Pain, 156(5), 825-836.
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3

Metabolic Profiling of Thymocytes in TBI

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Thymuses from untreated and SL-TBI treated mice were harvested and enzymatically digested and stained from flow cytometry analysis of thymocyte populations as above. Further analysis of mitochondrial bioenergetics were assessed using TMRE (Abcam, ab113852), MitoTracker™ Green FM (Invitrogen, M7514), MitoSOX™ Red Mitochondrial Superoxide Indicator (ThermoFisher, M36008), and Intracellular glutathione (GSH) Detection Assay Kit (Abcam, ab112132). Thymocytes were isolated from untreated and TBI-treated mice and intracellular pyruvate and lactate levels were measured by absorbance using Pyruvate Assay kit (Abcam, ab65342) or Lactate-Glo™ Assay (Promega, J5021). Thymocytes were isolated from untreated and TBI-treated mice and incubated in RPMI with 5 mM sodium pyruvate (Gibco, 11360070) for 3 h at 37 °C and stained for flow cytometry analysis and cl-caspase 1 levels. Cells were further incubated with 5 mM sodium pyruvate plus 200 μM α-ketobutyrate (Sigma-Aldrich) or 100 μM TEMPOL (Tocris, 3082) for 3 h and cells were prepared for flow cytometry analysis.
Kinsella S., Evandy C.A., Cooper K., Cardinale A., Iovino L., deRoos P., Hopwo K.S., Smith C.W., Granadier D., Sullivan L.B., Velardi E, & Dudakov J.A. (2023). Damage-induced pyroptosis drives endog thymic regeneration via induction of Foxn1 by purinergic receptor activation. bioRxiv.
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4

Aβ-induced Neurodegeneration Mitigants

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Coenzyme Q10 was generously provided by Kaneka Corporation. Human Aβ25–35 and tempol were purchased from Tocris. Human Aβ25–35 labelled with HiLyte Fluor 488 was obtained from Anaspec. The fluorescent probes, Fluo-4 AM, H2DCF-DA, MitoSOX-AM, MitoTracker Deep Red, Calcein-AM and Hoescht were obtained from LifeSicences. Alpha tocopherol was acquired from Sigma-Aldrich.
Durán-Prado M., Frontiñán J., Santiago-Mora R., Peinado J.R., Parrado-Fernández C., Gómez-Almagro M.V., Moreno M., López-Domínguez J.A., Villalba J.M, & Alcaín F.J. (2014). Coenzyme Q10 Protects Human Endothelial Cells from β-Amyloid Uptake and Oxidative Stress-Induced Injury. PLoS ONE, 9(10), e109223.
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5

Oxidative Stress Modulation in MG-63 Cells

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MG-63 cells were treated with indicated concentrations of 15d-PGJ2 (Cayman, MI, USA) for indicated time periods. Cells were incubated for 30 min with either an inhibitor of p38 MAPK (25 μM PD169316, Merck Biosciences, Darmstadt, Germany), N-acetyl-l-cysteine (NAC, 5 mM), pyrrolidine dithiocarbamate (PDTC, 1 mM), N-2-mercaptopropionylglycine (MPG, 5 mM) (Sigma–Aldrich, MO, USA), Tempol (1 mM, Tocris, MA, USA), PPARγ antagonist (20 μM T0070907), D-type prostanoid receptor (DP1) antagonist (100 nM MK0524), DP2 antagonist (1 μM CAY10471) (Cayman), l-buthionine-(S,R)-sulfoximine (BSO, 5 μM), GSH ethyl ester (10 mM, Sigma–Aldrich), LY294002 (10 μM) or Akt inhibitor (Akt-I, 10 μM) (Calbiochem, CA, USA) prior to 15d-PGJ2 treatment. Alternatively, cells were treated with 9,10-dihydro-15d-PGJ2 (dh-15d-PGJ2, 20 μM, Cayman) for indicated time periods.
Koyani C.N., Kitz K., Rossmann C., Bernhart E., Huber E., Trummer C., Windischhofer W., Sattler W, & Malle E. (2016). Activation of the MAPK/Akt/Nrf2-Egr1/HO-1-GCLc axis protects MG-63 osteosarcoma cells against 15d-PGJ2-mediated cell death. Biochemical pharmacology, 104, 29-41.
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6

Preparation of Morphine and Tempol Solutions

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Saline (vehicle) and morphine sulfate solution (50 mg/ml) were purchased from Hospira Inc (Lake Forest, IL, United States). Working dilutions of morphine (10 mg/ml) was prepared in sterile saline. Tempol was purchased from Tocris Bioscience (Minneapolis, MN, United States) and the stock solution (300 mg/ml) was prepared in sterile saline. Working dilutions (60 mg/ml and 100 mg/ml) were also prepared in sterile saline.
Baby S.M., Discala J.F., Gruber R., Getsy P.M., Cheng F., Damron D.S, & Lewis S.J. (2021). Tempol Reverses the Negative Effects of Morphine on Arterial Blood-Gas Chemistry and Tissue Oxygen Saturation in Freely-Moving Rats. Frontiers in Pharmacology, 12, 749084.
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7

Preparation of Stock Solutions for Pharmacological Agents

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Stock solutions of phenylephrine, acetylcholine and Y27632 were prepared in distilled water. Apocynin, cyclosporine H, LXA4, SQ 29548 and indomethacin were dissolved in ethanol; SC-560, NS-398 and tempol were dissolved in dimethyl sulfoxide and WRW4 was dissolved in 20% acetonitrile. All drugs were purchased from Sigma-Aldrich (Saint Louis, Missouri, USA) except for NS-398 and SC560 (Calbiochem, USA), WRW4 and tempol (Tocris bioscience, UK). The concentrations of the inhibitors and antagonists were selected based on previous studies [11 (link)-13 (link)]
Wenceslau C.F., McCarthy C.G., Szasz T, & Webb R.C. (2015). LIPOXIN A4 MEDIATES AORTIC CONTRACTION VIA RHOA/RHO KINASE, ENDOTHELIAL DYSFUNCTION AND REACTIVE OXYGEN SPECIES. Journal of vascular research, 51(6), 407-417.
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8

Transfection Reagents and Signaling Inhibitors

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Polyfect and Superfect transfection reagents were purchased from Qiagen, USA. AG1478 (N-(3-Chlorophenyl)-6,7-dimethoxy-4-quinazolinanine hydrochloride) and Tempol (4-hydroxy-2,2,6,6tetramethyl-piperidin-1-oxyl) were purchased from Tocris (UK). Epidermal growth factor (EGF) ligand, apocynin, catalase and all other reagents were purchased from Sigma Chemical Co (St Louis, USA).
Akhtar S., Al-Zaid B., El-Hashim A.Z., Chandrasekhar B., Attur S., Yousif M.H, & Benter I.F. (2015). Cationic Polyamidoamine Dendrimers as Modulators of EGFR Signaling In Vitro and In Vivo. PLoS ONE, 10(7), e0132215.
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9

Apoptosis Induction Assay with Flow Cytometry

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Unless otherwise noted, all materials were purchased from Sigma-Aldrich (St. Louis, MO). Cayman Chemical (Ann Arbor, MI) synthesized the TVX. Recombinant human TNF, z-VAD-fmk (ZVAD) and caspase 3 fluorometric assay kit were purchased from R&D Systems (Minneapolis, MN). Phosphate-buffered saline (PBS), high glucose Dulbecco’s Modified Eagles Medium (DMEM), Antibiotic-Antimycotic (ABAM), Lglutamine, and 0.25% trypsin-EDTA were purchased from Life Technologies (Carlsbad, CA). For flow cytometry experiments, Cell Staining Buffer was purchased from Biolegend (San Diego, CA), Perm/Wash Buffer from BD Biosciences (San Jose, CA), and Propidium Iodide/RNase Staining Solution from Cell Signaling Technology (Beverly, MA). U0126 was purchased from Calbiochem (San Diego, CA). KU55933 and Tempol were purchased from Tocris Bioscience (Minneapolis, MN). Cellular reactive oxygen species assay kit was purchased from Abcam (Cambridge, MA).
Beggs K.M., Maiuri A.R., Fullerton A.M., Poulsen K.L., Breier A.B., Ganey P.E, & Roth R.A. (2015). Trovafloxacin-induced Replication Stress Sensitizes HepG2 Cells to Tumor Necrosis Factor-alpha-induced Cytotoxicity Mediated by Extracellular Signal-regulated Kinase and Ataxia Telangiectasia and Rad3-related. Toxicology, 331, 35-46.
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10

Oxidative Stress Modulation in MG-63 Cells

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MG-63 cells were treated with indicated concentrations of 15d-PGJ2 (Cayman, MI, USA) for indicated time periods. Cells were incubated for 30 min with either an inhibitor of p38 MAPK (25 μM PD169316, Merck Biosciences, Darmstadt, Germany), N-acetyl-l-cysteine (NAC, 5 mM), pyrrolidine dithiocarbamate (PDTC, 1 mM), N-2-mercaptopropionylglycine (MPG, 5 mM) (Sigma–Aldrich, MO, USA), Tempol (1 mM, Tocris, MA, USA), PPARγ antagonist (20 μM T0070907), D-type prostanoid receptor (DP1) antagonist (100 nM MK0524), DP2 antagonist (1 μM CAY10471) (Cayman), l-buthionine-(S,R)-sulfoximine (BSO, 5 μM), GSH ethyl ester (10 mM, Sigma–Aldrich), LY294002 (10 μM) or Akt inhibitor (Akt-I, 10 μM) (Calbiochem, CA, USA) prior to 15d-PGJ2 treatment. Alternatively, cells were treated with 9,10-dihydro-15d-PGJ2 (dh-15d-PGJ2, 20 μM, Cayman) for indicated time periods.
Koyani C.N., Kitz K., Rossmann C., Bernhart E., Huber E., Trummer C., Windischhofer W., Sattler W, & Malle E. (2016). Activation of the MAPK/Akt/Nrf2-Egr1/HO-1-GCLc axis protects MG-63 osteosarcoma cells against 15d-PGJ2-mediated cell death. Biochemical pharmacology, 104, 29-41.
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