Cells were transfected and treated with indicated drugs for 24 hours and then collected. Hydroxyl radicals were detected using a Hydroxyl Radical Assay Kit (Jiancheng, Nanjing, China), ATP was measured using an ATP assay kit (Beyotime Biotechnology, Shanghai, China), cell oxygen consumption rate (OCR) was measured using Mito‐Xpress and pH‐Xtra kits (Luxcel Bioscience, Cork, Ireland), mitochondria extraction from cultured cells was performed (MP‐007, Inventbiotech), ACO2 activity was determined using an Aconitase Activity Assay Kit (MAK051, Sigma), and isocitrate and citrate concentrations were determined using an IsoCitrate Assay Kit (MAK319, Sigma) and a Citrate Assay Kit (MAK057, Sigma), respectively. All assays were performed according to the manufacturers' protocols.
Hydroxyl radical assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The Hydroxyl radical assay kit is a laboratory tool used to detect and quantify the presence of hydroxyl radicals in a sample. Hydroxyl radicals are highly reactive chemical species that play a role in various biological and environmental processes. The kit provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
Atp assay kit, by Beyotime (1 mentions)
Aconitase activity assay kit, by Merck Group (1 mentions)
Citrate assay kit, by Merck Group (1 mentions)
Isocitrate assay kit, by Merck Group (1 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Merck Group (1 mentions)
Vitamin c, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Bicinchoninic acid bca protein quantification kit, by Solarbio (1 mentions)
Cell counting kit 8 cck 8, by Keygen Biotech (1 mentions)
T aoc, by Nanjing Jiancheng (1 mentions)
Cell counting kit 8 cck 8 assay kit, by Beyotime (1 mentions)
H2o2 detection kit, by Nanjing Jiancheng (1 mentions)
Phosphate buffered saline (pbs), by Procell (1 mentions)
Ros assay kit, by Abcam (1 mentions)
Superoxide anion assay kit, by Nanjing Jiancheng (1 mentions)
Sodium tetrachloropalladate ii na2pdcl4, by Merck Group (1 mentions)
Nitric acid hno3, by Sinopharm (1 mentions)
Sod assay kit, by Dojindo Laboratories (1 mentions)
6 protocols using hydroxyl radical assay kit
1
Mitochondrial Bioenergetics Profiling
2
Antioxidant and Hepatoprotective Assays
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Vitamin C (VC) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Cell counting kit 8 (CCK-8) and caspase-3, caspase-8, and caspase-9 fluorescence metric assay kits were purchased from KeyGen BioTECH (Jiangsu, China). ALT, AST, T-AOC, SOD, DCF-DA cellular reactive oxygen species (ROS) detection assay kit, hydroxyl radical assay kit, and protein carbonyls assay kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The measurement kits for AChE and CAT activity were purchased from Comin Biotechnology (Suzhou, China). 8-hydroxy-2-deoxyguanosine (8-OHdG) ELISA kit was purchased from Wuhan ELISA Lab (Wuhan, China). The Annexin V-FITC and PI double staining assay kit was purchased from Vazyme Biotech (Nanjing, China). 4′,6-diamidino-2-phenylindole (DAPI) and the bicinchoninic acid (BCA) protein quantification kit were purchased from Solarbio (Beijing, China). Radioimmunoprecipitation assay (RIPA) cell lysis buffer was purchased from NCM Biotech (Suzhou, China).
3
Hydroxyl Radical Scavenging Assay of rTbprx-1
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The hydroxyl radical scavenging activity of rTbprx-1 was detected using theHydroxyl Radical Assay Kit(A018; Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Glutathione was used as the positive control experimental group; blank tubes, control tubes, and test tubes were set up to form three groups each for each reaction. After adding the reagent solution in a 37 °C water bath for 1 min, 2 mL of color developer prepared in advance was added and the mixture was protected from light for 10 min. The OD550 value was measured using an enzyme marker. The hydroxyl radical inhibition rate was calculated according to the instructions.
4
Antioxidant and Cytotoxicity Assays
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Sodium tetrachloropalladate (II) (Na2PdCl4, > 98%) was purchased from Sigma-Aldrich Trading Co., Ltd. (Shanghai, China). Insulin (bovine pancreatic, 27 u/mg), sodium hydroxide (NaOH), hydrochloric acid (HCl, 36–38%) and nitric acid (HNO3, 65–68%) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). A superoxide anion assay kit, hydroxyl radical assay kit and H2O2 detection kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). A Cell Counting Kit-8 (CCK-8) assay kit was obtained from Beyotime Institute of Biotechnology (Haimen, Jiangsu, China), and an ROS assay kit was purchased from Abcam Inc. (ab238535). PBS was obtained from Procell Life Science and Technology Co., Ltd. (Wuhan, China). Ultrapure water (18.2 MΩ cm− 1) was purified with a Milli-Q system.
5
Antioxidant Activity and Phenolic Profiling
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A total antioxidant activity assay was performed with the herbal extracts using a kit from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Determining the hydroxyl radical suppression of the herbal extracts was conducted according to the Fenton reaction principle. The procedure was performed using a hydroxyl radical assay kit, according to the manufacturer’s instructions (Nanjing Jiancheng Bioengineering Institute). Determination of the total phenolic content in the herbal extracts was measured using Lowry reagents (Sigma-Aldrich, St. Louis, MO, USA).
6
Antioxidant Activities of CeO2 Nanoparticles
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The SOD-mimicking activity of CeO 2 NPs was detected using a SOD assay kit (Dojindo). The reaction between WST-1 and O 2
•produced a water-soluble coloured product which could be detected using a microplate reader. While SOD inhibited the reduction reaction of O 2
•-by WST-1, SOD-mimicking activity could be calculated from the absorbance values of different groups.
The hydroxyl radical scavenging activity of CeO 2 NPs was investigated using a hydroxyl radical assay kit (Nanjing Jiancheng Bioengineering Institute). The Griess reagent reacted with • OH generated from the Fenton reaction of Fe 2+ and H 2 O 2 ; the absorbance value at 550 nm thus reflected the quantity of • OH present. Elimination of • OH was calculated utilizing the following equation: elimination ability (U mL -1 ) = (A c -A x )/(A s -A 0 ) × 8.824 mmol L -1 ; where A c denotes the control absorbance value, A x denotes the absorbance value of CeO 2 solutions at different concentrations, A s denotes the standard absorbance value, and A 0 denotes the blank absorbance value.
•produced a water-soluble coloured product which could be detected using a microplate reader. While SOD inhibited the reduction reaction of O 2
•-by WST-1, SOD-mimicking activity could be calculated from the absorbance values of different groups.
The hydroxyl radical scavenging activity of CeO 2 NPs was investigated using a hydroxyl radical assay kit (Nanjing Jiancheng Bioengineering Institute). The Griess reagent reacted with • OH generated from the Fenton reaction of Fe 2+ and H 2 O 2 ; the absorbance value at 550 nm thus reflected the quantity of • OH present. Elimination of • OH was calculated utilizing the following equation: elimination ability (U mL -1 ) = (A c -A x )/(A s -A 0 ) × 8.824 mmol L -1 ; where A c denotes the control absorbance value, A x denotes the absorbance value of CeO 2 solutions at different concentrations, A s denotes the standard absorbance value, and A 0 denotes the blank absorbance value.
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