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5 protocols using total sod activity detection kit

1

Investigating Liver Inflammation and Metabolism

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Vitamin C (Tianjin Kingyork, H12020392), Vitamin D3 (Beijing Solarbio Science & Technology, V8070), Mouse TNF-alpha ELISA Kit (RayBiotech, P06804), Mouse IL-6 ELISA Kit (RayBiotech, P08505), Mouse IL-1β ELISA Kit (Shanghai Tongwei Biotechnology, TW56961), Mouse insulin ELISA Kit (Shanghai Tongwei Biotechnology, TW085566), Mouse endotoxin (ET) ELISA Kit (Jiangsu Meimian, MM-0369M1), Total Cholestenone Content Assay Kit (Beijing Solarbio Science & Technology, BC 1985), Triglyceride Content Assay Kit (Beijing Solarbio Science & Technology, BC0625), Total SOD activity detection kit (Shanghai Beyotime Biotechnology, S0101S), Lipid oxidation detection kit (Shanghai Beyotime Biotechnology, S0131S), BCA protein concentration assay kit (Shanghai Beyotime Biotechnology, P0010), Oil red O (Sigma, O0625), Anti-FXR1 antibody (Abcam, ab155124), BSEP Monoclonal antibody (Proteintech Group, 67512-1-Ig), Anti-Fas antibody (Abcam, ab271016), Anti-ZO1 tight junction protein antibody (Abcam, ab221547), Anti-Occludin antibody (Abcam, ab222691), Anti-SLC10A2/ASBT antibody (Abcam, ab203205), TRIzol Reagent (Thermo Fisher, 15596026), cDNA synthesis kit (Beijing Tiangen Biotech, KR118), and SuperReal PreMix Plus (SYBR Green) (Beijing Tiangen Biotech, FP205) were used.
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2

Cytokine and Oxidative Stress Assays

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The levels of inflammatory factors tumour necrosis factor alpha (TNF-α), interleukin 1beta (IL-1β), IL-4, IL-5, and IL-13 in medium were determined by ELISA kit. The levels of ROS, ATP, catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in cells were also measured by ELISA kits. The kit details are as follows: Human TNF-α ELISA Kit (E-EL-H0109c; Elabscience, Wuhan, China), Human IL-1β ELISA Kit (E-EL-H0149c; Elabscience), Human IL-4 ELISA Kit (ml058093; mlbio, Beijing, China), Human IL-5 ELISA Kit (ml058095; mlbio), Human IL-13 ELISA Kit (ml063719; mlbio), Reactive Oxygen Species detection Reagent kit (S0033M; Beyotime, Beijing, China), ATP Detection Kit (S0026; Beyotime), Human Catalase ELISA Kit (EH0643; Wuhan Fine Biotech, Wuhan, China), Total SOD Activity Detection Kit (S0101S; Beyotime), and Glutathione Peroxidase Detection Kit (S0056; Beyotime). All procedures were carried out in accordance with the manufacturers’ instructions. Each experiment was repeated with at least three independent experiments.
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3

Measurement of Intracellular SOD Activity

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The intracellular SOD activity was measured with a total SOD activity detection kit (Beyotime Biotechnology, S0101, Shanghai, China), according to the manufacturer’s instructions. After drug treatment, the cells were washed once with PBS and collected by centrifugation. The cells were fully lysed in SOD sample preparation solution, and the supernatants were collected at 12,000 × g at 4°C for 5 min. The absorbance was detected at 450 nm and 600 nm (reference wavelength) with a multimode microplate reader, and the SOD activity was calculated.
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4

Determination of Superoxide Dismutase Activity

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The superoxide dismutase (SOD) activity was determined using the total SOD activity detection kit according to the WST-8 method (Beyotime, Shanghai, China) [32 (link)]. Twenty microliters of appropriately diluted samples were added to 160 μL WST-8/enzyme working solution and 20 μL reaction starting solution and incubated at 37 °C for 30 min; we then determined the absorbance value at 450 nm. Blank 1 was 20 μL SOD test buffer added to 160 μL WST-8/enzyme working solution and 20 μL reaction starting solution; Blank 2 was 40 μL SOD test buffer added to 160 μL WST-8/enzyme working solution; and Blank 3 was 20 μL of appropriately diluted sample added to 20 μL SOD test buffer and 160 μL WST-8/enzyme working solution.
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5

Measuring Ovarian SOD Activity

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Mouse ovaries and ovarian tissue homogenates were obtained according to the Tunnel kit and total SOD activity detection kit (S0101S, Beyotime, China), respectively.
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