After centrifugation, the supernatant fluid of BALF was collected for SOD and NO detection. 20 μl and 100 μL supernatant was used to detect the levels of SOD and NO in BALF, respectively. SOD level was analyzed by SOD assay kit (WST-1 method) in accordance to the manufacturer’s directions (Jiancheng Biotechnology, Nanjing, China, A001-3). That was an activity test and the substrate was water soluble tetrazolium salt (WST), which could be easily reduced to purple formazan. Then, the OD value (450 nm) was detected with a microplate reader. Activity of SOD was calculated using the following formula: SOD activity (U/mL)=SOD inhibition rate/50% x (reaction volume/dilution ratio) x dilution ratio of samples before detection. NO level was detected using NO assay kit (microwell plate method) (Jiancheng Biotechnology, Nanjing, China, A013-2) and evaluated by the following formula: NO level (μmol/L)=(samples OD value – blank OD value)/(standard OD value – blank OD value) x standard concentration (20 μm) x dilution ratio.
Sod assay kit wst 1 method
Manufactured by Nanjing Jiancheng
Sourced in China
The SOD assay kit (WST-1 method) is a laboratory equipment designed to measure the activity of superoxide dismutase (SOD) in biological samples. The kit utilizes the WST-1 (water-soluble tetrazolium salt) method to quantify SOD levels. It provides a simple, rapid, and reliable way to assess the antioxidant capacity of various samples.
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Lab products found in correlation
No assay kit, by Nanjing Jiancheng (1 mentions)
Nitric oxide assay kit, by Nanjing Jiancheng (1 mentions)
Mda assay kit tba method, by Nanjing Jiancheng (1 mentions)
Enspire multimode plate reader, by PerkinElmer (1 mentions)
Atp assay kit, by Beyotime (1 mentions)
Cat assay kit, by Beyotime (1 mentions)
Trehalose content detection kit, by Nanjing Jiancheng (1 mentions)
Bca protein assay kit, by Dingguo (1 mentions)
Mda assay kit, by Nanjing Jiancheng (1 mentions)
T aoc assay kit, by Nanjing Jiancheng (1 mentions)
Gsh px assay kit, by Nanjing Jiancheng (1 mentions)
Cat assay kit, by Nanjing Jiancheng (1 mentions)
8 protocols using sod assay kit wst 1 method
1
Quantifying SOD and NO in BALF
2
Evaluating Oxidative Stress Markers in Ischemic Rat Brains
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Six rats from each of above three groups were randomly selected on the first, third and seventh day after model establishment. The animals were killed, and the brains were rapidly removed. The infarct‐side cortexes were rapidly frozen in liquid nitrogen and then stored at −80°C. The cortex tissue samples were homogenized in cold saline to make 10% homogenates. They were then centrifuged at 1248 g, and the supernatants were collected and stored at −80°C. Nitric oxide and MDA contents and iNOS and SOD activities were measured according to the manufacturer's instructions using the nitric oxide assay kit (A013‐2; Nanjing Jiancheng Biology Engineering Institute, Nanjing, China), MDA assay kit (TBA method) (A003‐1; Nanjing Jiancheng Biology Engineering Institute), NOS typed assay kit (Colormetric method) (A014‐1‐1; Nanjing Jiancheng Biology Engineering Institute), and SOD assay kit (WST‐1 method) (A001‐3; Nanjing Jiancheng Biology Engineering Institute), respectively.
3
Evaluating Superoxide Dismutase-like Activity of Cu-based Nanoparticles
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The SOD-like activity of Cu5.4O USNPs was determined by formazan formation using a SOD assay kit (WST-1 method) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China)19 (link). Briefly, O2− was generated through the oxidation of xanthine by xanthine oxidase (XO), which can convert the WST-1 into WST-1 formazan with a characteristic absorption at 450 nm. The formazan concentration was determined at 450 nm using a multiple plate reader. The SOD-like activity of Cu5.4O USNPs was further confirmed with EPR spectroscopy. Briefly, a series of samples containing xanthine (5 mM) and xanthine oxidase (0.5 U mL−1) in 10 mM PBS and incubated for 10 min at 37 °C. Different Cu5.4O USNPs solutions (0, 20, 100, 1000 ng mL−1) were added and then the EPR signals were recorded immediately.
4
Evaluating Superoxide Dismutase Activity
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To assess superoxide dismutase (SOD) activity, wild-type worms were cultured in NGM plates containing 160 mg/L MDHB or not for 6 days. Animals on the sixth day of adulthood were collected from different plates with M9 buffer and washed thrice. Afterward, the harvested animals were suspended in M9 buffer and homogenized on ice [46 (link)]. The EnSpire Multimode Plate Reader (PerkinElmer, Waltham, MA, USA) was used to evaluate SOD activity according to an SOD assay kit (WST-1 method) (Nanjing Jiancheng, Nanjing, China).
5
Cellular Antioxidant and Energy Profiles
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After collection, the cells were washed and re-suspended in the PBS buffer. Then the SOD was analyzed by using the superoxide dismutase (SOD) assay kit (WST-1 method) (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). The catalase (CAT) activity was detected by using the CAT assay kit (Beyotime Biotechnology, Shanghai, China)). The ATP content was measured using an ATP assay kit (Beyotime Biotechnology, Shanghai, China). The protein concentration was measured using the bicinchonininc acid (BCA) protein assay kit (Dingguo Changsheng Biotechnology Co., LTD, Beijing, China) to adjust enzyme activities and ATP content. The intracellular trehalose was extracted and determined by a trehalose content detection kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
6
Antioxidant Capacity Assessment Protocol
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Total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT), and malondialdehyde (MDA) were determined to assess antioxidant capacity. T-AOC assay kit, SOD assay kit (WST-1 method), GSH-PX assay kit, CAT assay kit (visible light), and MDA assay kit (TBA method) were bought from Nanjing Jiancheng Bioengineering Institute (Nanjing, China), and the inspection process was carried out in accordance with the instructions.
7
Antioxidant Enzyme Assays in Samples
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CAT assay kit (micro-method) (BC0205) and MDA assay kit (micro-method) (BC0025) were purchased from Beijing Solarbio Science & Technology Co., Ltd (Beijing, China), while SOD assay kit (WST-1 method) (A001-3) was obtained form Nanjing Jiancheng Bioengineering Institute (Nanjing, China). After operating following the instructions, the date of results was collected and analyzed.
8
Spectrophotometric Determination of SOD
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The activity of total SOD (EC 1.15.1.1) was determined using a SOD assay kit (WST-1 method) purchased from the Nanjing Jiancheng Bioengineering Institute (WST-1 method). A 0.1 g of fresh sample was ground in liquid nitrogen, homogenized in a 0.9 mL ice bath by adding 10 mM phosphate buffer (pH 7.4), and then the mixture was centrifuged at 8000 × g for 10 minutes at 4 °C. SOD activity was expressed as U g−1, where U = 0.01ΔOD450 min−1. And one unit (U) of SOD activity is defined as the amount of enzyme corresponding to a SOD inhibition rate of 50%.
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