Superoxide dismutase

Manufactured by Cayman Chemical

Sourced in United States

Superoxide dismutase is an enzyme that catalyzes the conversion of superoxide radicals into oxygen and hydrogen peroxide. It plays a crucial role in the body's antioxidant defense system by neutralizing reactive oxygen species.

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7 protocols using superoxide dismutase

Postmortem Lung Perfusion and Analysis

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Postmortem, the lungs were removed and perfused with saline. The left lung was isolated, occluded, and stored at –80°C until use in biochemical analysis, whereas the right lung was fixed (4% formalin) for histological analysis (19 (link)).
Frozen lung samples were used for measurements of interleukin (IL)–8, IL-1B, and tumor necrosis factor-α concentration using specific enzyme-linked immunosorbent assay kits (Abnova, Tapei City, Taiwan). Enzyme activities, catalase (19 (link), 21 (link)), and superoxide dismutase were tested (Cayman Chemical, Ann Arbor, MI), whereas protein concentrations were determined by the Bradford method (22 (link)).
Formalin-fixed tissue was sectioned (5-µm thick), stained, and analyzed with light microscopy. Lung injury was scored by a pathologist blinded to treatment allocation. Pathologic signs of lung injury (atelectasis, alveolar and interstitial inflammation, alveolar and interstitial hemorrhage, edema, and necrosis) were each scored on a 0- to four-point scale: 0 corresponding to no injury; 1, 2, and 3 to injury to 25%, 50%, and 75% of the field; and 4 to injury across the field (20 (link), 23 (link)). Additionally, all seven injury scores were summed to obtain a mean total lung injury score for each group, ranging from 0 to 28, values higher than 12 corresponding to quite severe lung injury (19 (link)).

Rey-Santano C., Mielgo V.E., Gomez-Solaetxe M.A., Bianco F., Salomone F, & Loureiro B. (2020). Nebulized Poractant Alfa Reduces the Risk of Respiratory Failure at 72 Hours in Spontaneously Breathing Surfactant-Deficient Newborn Piglets. Critical Care Medicine, 48(6), e523-e531.

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Neuroprotective Effects of Isoquercitrin in PC12 Cells

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PC12 cells were purchased from ATCC (#CRL-1721.1 PC12 ADH, RattusNorvegicus). 6-hydroxydopamine, isoquercitrin, poly-L-Lysine, MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide),3,4 – dihydroxy-L-phenylalanine (Levodopa) and dimethyl sulphoxide (DMSO) were purchased from Sigma-Aldrich (USA). Dulbecco’s modified eagle’s medium (DMEM), pen-strep, horse serum, and fetal bovine serum was purchased from GIBCO Inc. (USA). Kits for glutathione peroxidase, superoxide dismutase, catalase, thiobarbiturate assay, and glutathione were purchased from the Cayman Chemical Company (USA).

Magalingam K.B., Radhakrishnan A, & Haleagrahara N. (2014). Protective effects of flavonol isoquercitrin, against 6-hydroxy dopamine (6-OHDA) - induced toxicity in PC12 cells. BMC Research Notes, 7, 49.

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Synthesis and Evaluation of NOSH-Aspirin

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NOSH-aspirin (NBS-1120) 4-(3-thioxo-3H-1,2-dithiol-5-yl)phenyl 2-((4-(nitrooxy)butanoyl)oxy), benzoate was synthesized as described previously [9 (link)] and was a gift from Avicenna Pharmaceuticals Inc, (New York, NY). The structural components of the NOSH-aspirin are shown in Figure 1. Lipopolysaccharide (LPS) from E. coli, aspirin, and carrageenan were purchased from Sigma (St. Louis, MO, USA). The kits used for determination of PGE2, lipid peroxidation, and superoxide dismutase were purchased from Cayman Chemical (Ann Arbor, MI).

Kodela R., Chattopadhyay M., Velázquez-Martínez C.A, & Kashfi K. (2015). NOSH-aspirin (NBS-1120), a novel nitric oxide- and hydrogen sulfide-releasing hybrid has enhanced chemo-preventive properties compared to aspirin, is gastrointestinal safe with all the classic therapeutic indications. Biochemical pharmacology, 98(4), 564-572.

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Oxidative Stress Evaluation in Plasma

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Superoxide dismutase (Cayman Chemical, Item No. 706002) and total antioxidant power (Oxford Biomedical Research, #TA02) were measured with commercial assays according to manufacturers’ instructions on plasma samples. The SOD assay kit used a tetrazolium salt to detect the amount of superoxide radicals that were generated by xanthine oxidase and hypoxanthine. The SOD kit measured all three isoforms: Cu/Zn, Mn, and FeSOD. One unit of SOD in the sample was defined as the amount of enzyme needed to dismutate 50% of a superoxide radical. Sample concentrations were compared against a bovine erythrocyte SOD (Cu/Zn) standard and interpolated from a standard curve. Total antioxidant power was determined based on the ability of the sample to reduce Cu⁺² to Cu⁺¹ compared to a trolox standard.

Witkop J.J., Vertigan T., Reynolds A., Duffy L., Barati B., Jerome S, & Dunlap K. (2021). Sled dogs as a model for PM2.5 exposure from wildfires in Alaska. Environment international, 156, 106767.

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Evaluation of Antioxidant Potential in Kidney Cells

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Extract of SG was a generous gift from Ginseng Science Inc. (Seoul, Korea). Silica gel (230–400 mesh) was obtained from Merck (Darmstadt, Germany). Solvents for extraction and column chromatography were purchased from Duksan Pure Chemicals (Ansan, Korea). Diaion HP-20 was obtained from Mitsubishi Chemical Industries (Tokyo, Japan). Dulbecco's Modified Eagle Medium supplemented with Nutrient Mixture F-12 (DMEM/F-12), fetal bovine serum, and antibiotics (10,000 units/mL of penicillin G and 10,000 μg/mL streptomycin sulfate) for LLC-PK1 culture were purchased from Invitrogen (Carlsbad, CA, USA). For reactive oxygen species (ROS) measurement, 2′,7′-dichlorofluorescin diacetate (DCFH-DA) was obtained from Invitrogen. Diagnostic kits for measurement of blood urea nitrogen (BUN) and creatinine were purchased from Asan Pharmaceuticals (Seoul, Korea). Assay kits for catalase, glutathione reductase, and superoxide dismutase was purchased from Cayman (Ann Arbor, MI, USA). A Bradford Protein Assay kit (Thermo Scientific, Rockford, IL, USA) was used for determination of proteins in cell or kidney homogenates. Cisplatin, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), HPLC-grade solvents, and other reagents were purchased from Sigma–Aldrich (St Louis, MO, USA).

Baek S.H., Shin B.K., Kim N.J., Chang S.Y, & Park J.H. (2016). Protective effect of ginsenosides Rk3 and Rh4 on cisplatin-induced acute kidney injury in vitro and in vivo. Journal of Ginseng Research, 41(3), 233-239.

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Superoxide Dismutase Activity Assay

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Superoxide dismutase (SOD) assay kit (Cayman Chemical Company, Michigan, USA) is based on the conversion of a tetrazolium salt to formazan by superoxide radicals generated in xanthine/xanthinoxidase system. The current method measures the activities of all three SOD types. One unit of enzyme is defined as the amount of enzyme needed to exhibit 50% dismutation of the superoxide radical. Absorbances were monitored at 450 nm by HT BioTek Synergy (BioTek Instruments, USA) microplate plate reader.

Gal A.F., Stan L., Tăbăran F., Rugină D., Cătoi A.F, & Andrei S. (2020). Chemopreventive Effects of Propolis in the MNU-Induced Rat Mammary Tumor Model. Oxidative Medicine and Cellular Longevity, 2020, 4014838.

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Oxidative Stress Markers in Adipose Tissue

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Superoxide dismutase (catalog no. 706002; Cayman Chemical, Ann Arbor, MI), malondialdehyde (MDA; catalog no. 10009055; Cayman Chemical), GSH (catalog no. NWK-GSH01; Northwest Life Science Specialties, Vancouver, WA), and ROS (catalog no. STA-347, Cell Biolabs, San Diego, CA) were analyzed in subcutaneous adipose tissue using commercial kits. Concentration of total protein of the adipose samples was measured using the BCA assay kit (catalog no. 23227; Thermo Fisher Scientific).

Liang Y., Batistel F., Parys C, & Loor J.J. (2019). Glutathione metabolism and nuclear factor erythroid 2-like 2 (NFE2L2)-related proteins in adipose tissue are altered by supply of ethyl-cellulose rumen-protected methionine in peripartal Holstein cows. Journal of dairy science, 102(6).

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