S0058
Manufactured by Beyotime
Sourced in China
The S0058 is a laboratory equipment used for general purposes. It measures 12 x 8 x 6 inches and weighs approximately 5 pounds.
Automatically generated - may contain errors
Lab products found in correlation
S0101, by Beyotime (3 mentions)
S0051, by Beyotime (2 mentions)
S0103, by Beyotime (2 mentions)
Synergy ht multi mode microplate reader, by Agilent Technologies (1 mentions)
Micro total mercapto assay kit, by Solarbio (1 mentions)
S0053, by Beyotime (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Fv500, by Olympus (1 mentions)
Bc0025, by Solarbio (1 mentions)
Gsh px, by Beyotime (1 mentions)
7 protocols using s0058
1
Antioxidant Enzyme Activity Determination
2
Glutathione Peroxidase Activity Assay
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The methods of protein extraction and concentration determination were described by Western blotting assay. The enzyme activity of total GPx was measured using a commercial test kit (Beyotime, S0058) following the manufacturer’s instructions. The decrease in nicotinamide adenine dinucleotide phosphate measured at 340 nm was linearly correlated with enzyme activity using a PowerWave XS spectrophotometer.
3
Assessing Glutathione Redox Status
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After different treatment, proteins were harvested to determine the activities of glutathione peroxidase (GPX) and GR and assess the content of reduced glutathione (GSH) as well as GSH/oxidized glutathione (GSSG) ratio in accordance with the responding assay kit (Beyotime, #S0058, #S0055 or #S0053).
4
Antioxidant Enzyme Assays in Cells
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ELISA detection kits for total superoxide dismutase (SOD, S0103, Beyotime Institute of Biotechnology, Nantong, China), total glutathione peroxidase (GPx, S0058, Beyotime Institute of Biotechnology), and malondialdehyde (MDA, S0131S, Beyotime Institute of Biotechnology) were used for measurements. Assays were performed following the manufacturer’s instructions. The brain tissues were isolated on day 7 and placed in 0.9% cold saline. Samples were homogenized and centrifuged at 3000 r/minute for 15 minutes. The supernatant was collected for ELISA, and samples were analyzed using a Synergy™ HT multimode microplate reader (Biotek, Shanghai, China).
To measure MDA, SOD and GPx activities in SH-SY5Y cells, cells were seeded at 1 × 105 cells/well in 96-well plates for 24 hours, followed by co-treatment with 100 μM hemin at various concentrations (1, 10 and 100 nM) of WFA or ferrostatin-1 (fer-1, 10 μM) for 24 hours. The cells were collected using a rubber scraper and centrifuged at 1000 × g for 10 minutes at 4°C. Cell pellets were lysed in 500 μL of 5% (w/v) metaphosphoric acid. Cellular lysate was centrifuged at 13,000 × g for 5 minutes at 4°C. Supernatants were collected for measurement of GSH-Px, SOD and MDA activities using a Synergy™ HT multimode microplate reader (Biotek).
To measure MDA, SOD and GPx activities in SH-SY5Y cells, cells were seeded at 1 × 105 cells/well in 96-well plates for 24 hours, followed by co-treatment with 100 μM hemin at various concentrations (1, 10 and 100 nM) of WFA or ferrostatin-1 (fer-1, 10 μM) for 24 hours. The cells were collected using a rubber scraper and centrifuged at 1000 × g for 10 minutes at 4°C. Cell pellets were lysed in 500 μL of 5% (w/v) metaphosphoric acid. Cellular lysate was centrifuged at 13,000 × g for 5 minutes at 4°C. Supernatants were collected for measurement of GSH-Px, SOD and MDA activities using a Synergy™ HT multimode microplate reader (Biotek).
5
Antioxidant Enzyme Activities Quantification
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After stromal cells were lysed and centrifuged, supernatants were collected for analyzing the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) by the corresponding assay kit (Beyotime, S0103, S0051, or S0058). For SOD activity, supernatants were incubated with WST-8/enzyme solution concomitant with an addition of reaction-started working solution, and then, the absorbance was measured at 450 nm to calculate SOD activity in accordance with the corresponding formula. For CAT activity, supernatants were mixed with 5 mM H2O2 for 5 min followed by a supplementation of substrate solution, and then, the absorbance was assessed at 520 nm to calculate CAT activity according to standard curve. For GPX activity, supernatants were mixed with working solution along with the addition of peroxide reagent and then determined the absorbance at 340 nm to calculate GPX activity in the light of the corresponding formula.
6
Oxidative Stress Assessment in Cells
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After the mice or cells were treated, cells or liver tissues were lysed in ice-cold RIPA Lysis buffer (P0013B, Beyotime) containing 1 mM phenylmethylsulfonyl fluoride (PMSF). The lysate was collected and put into EP tube, and the protein concentration was detected by BCA reagent method (P0010, Beyotime). Total sulfhydryl groups (TSH) in the cells or liver tissues was analyzed with the Micro Total Mercapto Assay Kit (BC1375, Solarbio, Beijing, China) according to the manufacturer’s protocol. The levels of MDA (SO0131, Beyotime), GSH/GSSG (S0053, Beyotime), and activities of SOD (S0101, Beyotime), GSH-Px (S0058, Beyotime), CAT (S0051, Beyotime) were measured with commercial assay kits according to the manufacturers’ protocols.
7
Quantifying Oxidative Stress in Hepatocytes
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The ROS content in hepatocytes was determined with dichlorofluorescein diacetate (S0033S; Beyotime Biotechnology Inc.). Briefly, hepatocytes were incubated with dichlorofluorescein diacetate for 30 min at 37°C. The fluorescence intensity of cells was measured by flow cytometry (CytoFLEX, Beckman Coulter) and laser confocal microscopy (FV500, Olympus). Superoxide dismutase (SOD; S0101; Beyotime Biotechnology Inc.) and glutathione peroxidase (GSH-Px; S0058; Beyotime Biotechnology Inc.) activities were measured using commercial kits. Malondialdehyde (MDA) content was measured using spectrophotometric diagnostic kits (BC0025; Solarbio Technology Co., Ltd).
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