Tempone h

Manufactured by Enzo Life Sciences

Sourced in United States

TEMPONE-H is an electron paramagnetic resonance (EPR) spin probe used for the detection and quantification of oxidative stress in biological samples. It is a nitroxide radical that exhibits characteristic EPR signals, which can be used to measure the presence and levels of reactive oxygen species.

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Lab products found in correlation

Hydrogen peroxide peroxidase assay kit, by Thermo Fisher Scientific (2 mentions) Esr spectroscope, by Bruker (2 mentions) Nitrate nitrite colorimetric assay kit, by Cayman Chemical (2 mentions) 5 5 dimethyl 1 pyrroline n oxide dmpo, by Dojindo Laboratories (1 mentions) Griess reagent kit, by Beyotime (1 mentions) Esr spectroscopy, by Bruker (1 mentions) 5 5 dimethyl 1 pyrroline n oxide, by Dojindo Laboratories (1 mentions) Sodium azide, by Merck Group (1 mentions) Carboxy ptio, by Merck Group (1 mentions) Mannitol, by Merck Group (1 mentions) Hydrogen peroxide assay kit, by Beyotime (1 mentions) N methyl d glucaminedithiocarbamate mgd, by Dojindo Laboratories (1 mentions) S0038, by Beyotime (1 mentions) Esr spectrometer, by Bruker (1 mentions) Nitrite nitrate colorimetric assay kit, by Beyotime (1 mentions) Varioskan flash, by Thermo Fisher Scientific (1 mentions)

6 protocols using tempone h

Measuring Reactive Oxygen and Nitrogen Species

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The concentrations of H₂O₂ and NO₂⁻/NO₃⁻ in the water were measured using a hydrogen peroxide/peroxidase assay kit (Thermo Fisher Scientific) and a nitrite/nitrate colorimetric assay kit (Cayman), respectively. ˙OH, ¹O₂, ˙NO, O₂˙⁻, ˙NO₂, and ONOO⁻ were measured using an electron spin resonance (ESR) spectroscope (Bruker) with relevant spin traps (29 (link)). The spin traps were 100 mM 5,5-dimethyl-1-pyrroline N-oxide (DMPO; Dojindo) for trapping ˙OH, 5 mM N-(dithiocarbamoyl)-N-methyl-d-glucamine (MGD; Dojindo) for trapping ˙NO, 10 mM 2,2,6,6-tetramethylpiperidine (TEMP; TCI) for trapping ¹O₂, and 10 mM 1-hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine (TEMPONE-H; Enzo) for trapping O₂˙⁻, ˙NO₂, and ONOO⁻.

Guo L., Xu R., Gou L., Liu Z., Zhao Y., Liu D., Zhang L., Chen H, & Kong M.G. (2018). Mechanism of Virus Inactivation by Cold Atmospheric-Pressure Plasma and Plasma-Activated Water. Applied and Environmental Microbiology, 84(17), e00726-18.

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Quantification of Reactive Species

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Here, aqueous H₂O₂ was measured using the Amplex Red Hydrogen Peroxide/Peroxidase Assay kits (Beyotime, Shanghai, China) according to the manufacturer’s instructions. Aqueous NO₂⁻ and NO₃⁻ were measured using the Griess reagent kit (Beyotime, Shanghai, China). Aqueous OH• was detected using the DMPO (5,5-dimethyl-1-pyrrolineN-oxide, Dojindo, 5 mM) by an electron spin resonance (ESR) spectrometer (BrukerBioSpin GmbH, EMX, Ettlingen, Germany). The contents of aqueous O₂⁻/ONOO⁻/ONOOH were measured using the TEMPONE-H (1-hydroxy-2,2,6,6-tetramethyl-4-oxo-piper-idine, Enzo, 5 mM). Detailed ROS and RNS detection methods are outlined in our previous paper [45 (link)]. PAS sampling for H₂O₂, NO₂⁻ and NO₃⁻ measurement were completed within 5 min after plasma irradiation due to other operations. The DMPO (or TEMPONE-H) were added into the saline before plasma irradiation, and also within 5 min after irradiation the PAS sampling (containing DMPO or TEMPONE-H) was analyzed by ESR. All experiments were performed three times (n = 3).

Zhang H., Zhang J., Guo B., Chen H., Xu D, & Kong M.G. (2021). The Antitumor Effects of Plasma-Activated Saline on Muscle-Invasive Bladder Cancer Cells In Vitro and In Vivo Demonstrate Its Feasibility as a Potential Therapeutic Approach. Cancers, 13(5), 1042.

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Quantifying Reactive Species in Saline

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The concentrations of H₂O₂ and

{NO}_{2}^{-}

{NO}_{3}^{-}

in 0.9% NaCl were measured using hydrogen peroxide/peroxidase assay kit (Thermo Fisher Scientific) and nitrite/nitrate colorimetric assay kit (Cayman), respectively. ^•OH, ¹O₂, ^•NO,

O_{2}^{• -}

, ^•NO₂, and ONOO⁻ were measured using an electron spin-resonance (ESR) spectroscopy (Bruker) together with relevant spin traps. The latter include: 100 mM DMPO (5,5-Dimethyl-1-pyrroline N-oxide, Dojindo), 5 mM MGD (N-(Dithiocarbamoyl)-N-methyl-D-glucamine, Dojindo), 10 mM TEMP (2,2,6,6-Tetramethylpiperidine, TCI), and 10 mM TEMPONE-H (1-Hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine, Enzo).

Guo L., Xu R., Zhao Y., Liu D., Liu Z., Wang X., Chen H, & Kong M.G. (2018). Gas Plasma Pre-treatment Increases Antibiotic Sensitivity and Persister Eradication in Methicillin-Resistant Staphylococcus aureus. Frontiers in Microbiology, 9, 537.

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Bio-Gel Scavengers and Reactive Species

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Three model bio‐gels were HEC (ZRNCUN), Carbomer 940 (Carbomer, ZRNCUN), and AVC (ZRNCUN). The scavengers used in this study were mannitol (Sigma–Aldrich), sodium azide (Sigma–Aldrich), tiron (Acros Organics), ebselen (Biochempartner), and carboxy‐PTIO (Sigma–Aldrich). Reactive species detection reagents were disodium terephthalate (TPT, Aladdin), N‐(dithiocarbamoyl)‐N‐methyl‐D‐glucamine (MGD, Dojindo), TEMP (Aldrich), and TEMPONE‐H (Enzo). The hydrogen peroxide assay kit and the nitrate/nitrite colorimetric kit were from Beyotime Biotech.

Chen J., Wang Z., Sun J., Zhou R., Guo L., Zhang H., Liu D., Rong M, & Ostrikov K.(. (2023). Plasma‐Activated Hydrogels for Microbial Disinfection. Advanced Science, 10(14), 2207407.

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Quantifying CAP-generated Reactive Species

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An electron spin resonance (ESR) spectrometer (Bruker, Germany) and the following spin traps: 5,5-dimethyl-1-pyrroline N-oxide (DMPO, Dojindo Laboratories) were used. Long-lived ROS produced by CAP_PZ2 in PBS were measured using a microplate reader with a hydrogen peroxide assay kit (S0038, Beyotime Biotechnology, China) for H₂O₂. The Griess Reagent Nitrate/Nitrite kit was used for nitrite (NO₂⁻) and nitride (NO₃⁻). Short-lived ROS were measured including OH^•, N-(dithiocarboxy) sarcosine (DTCS) and N-methyl-d-glucamine dithiocarbamate (MGD; Dojindo Laboratories, Japan) for NO^•, 2,2,6,6-tetramethylpiperidine (TEMP; Sigma-Aldrich, St. Louis, MO, USA) for ¹O₂, and 1-hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine (TEMPONE-H; Enzo Biochem, Farmingdale, NY, USA) for O₂^•– and ONOO⁻. Spin trap concentrations were chosen based on dosing experiments. For 30s or 60 s CAP_PZ2 treatment, 5 mM TEMPONE-H was found to be adequate for trapping ONOO⁻ and O₂^•–. For TEMP and DMPO, 30 mM and 50 mM were chosen, respectively. Spin traps were added to PBS before and after CAP_PZ2 treatment for measurement of ROS produced during the treatment and residual ROS post-treatment, respectively.

Wang Y., Mang X., Li D., Wang Z., Chen Y., Cai Z, & Tan F. (2023). Cold atmospheric plasma sensitizes head and neck cancer to chemotherapy and immune checkpoint blockade therapy. Redox Biology, 69, 102991.

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Quantifying Reactive Species in PAA-Activated Sprays

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The water spray after the PAA treatment was recovered to determine the reactive species in the PAA-activated spray. The concentrations of H₂O₂ and NO₂⁻/NO₃⁻ in the recovered water were measured by using a hydrogen peroxide/peroxidase assay kit (AAT Bioquest) and nitrite/nitrate colorimetric assay kit (Beyotime Biotechnology), respectively, as previously reported [19] (link). The ^•OH and ¹O₂ were measured by using fluorescent probes of terephthalic acid disodium (TPT, Sigma), trans-1-(2′-methoxyvinyl)pyrene (tMVP, J&K Scientific) at final concentrations of 3 mM and 20 μM, respectively. After incubation, the fluorescence intensities were detected by using a microplate reader (Thermo Scientific Varioskan Flash) at the corresponding excitation and emission wavelengths (TPT: 310/425 nm; tMVP: 405/460 nm). The total concentrations of O₂⁻ and ONOO⁻ were determined using the spin trap 1-Hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine hydrochloride (TEMPONE-H, Enzo Life Sciences) at final concentrations of 1 mM and the spin adducts 4-oxo-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPONE) were detected using an electron spin resonance (ESR) spectroscope (Bruker).

Guo L., Zhao P., Jia Y., Li T., Huang L., Wang Z., Liu D., Hou Z., Zhao Y., Zhang L., Li H., Kong Y., Li J., Wang X, & Rong M. (2022). Efficient inactivation of the contamination with pathogenic microorganisms by a combination of water spray and plasma-activated air. Journal of Hazardous Materials, 446, 130686.

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