The accumulation level of H2O2 was observed using DAB as described previously (Zhu et al., 2020 (link)). Endogenous H2O2 concentrations were determined according to the protocol described by Chen et al. (2019 (link)) using a hydrogen peroxide assay kit (Solarbio) (Chen et al., 2019 (link)). Qualitative test of O2•− was observed using NBT with a similar procedure to that described previously (Zhu et al., 2020 (link)). The content of O2•− in N. benthamiana leaves was measured according to previously methods as described by Cai et al. (2018 (link)) using a Micro Superoxide Anion Assay Kit (Solarbio) (Cai et al., 2018 (link)).
Micro superoxide anion assay kit
Manufactured by Solarbio
Sourced in China
The Micro Superoxide Anion Assay Kit is a laboratory product designed to measure the concentration of superoxide anions (O2-) in biological samples. The kit provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
Hydrogen peroxide assay kit, by Solarbio (1 mentions)
Hydrogen peroxide assay kit, by Beyotime (1 mentions)
Lipid peroxidation mda assay kit, by Beyotime (1 mentions)
Superoxide dismutase sod activity detection kit, by Solarbio (1 mentions)
Catalase cat activity assay kit, by Solarbio (1 mentions)
Ascorbate peroxidase apx activity assay kit, by Solarbio (1 mentions)
5 protocols using micro superoxide anion assay kit
1
Quantification of H2O2 and O2•− in Plants
2
Peanut Seed Germination Stress Response
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The two varieties of peanut seeds were immersed in distilled water for 4 h, transferred to petri dishes at 2°C for 72 h, and then returned to normal temperature (28°C) for additional 3 days. The controls were continuously germinated at 28°C for 3 days. The germination rates were measured every day.
Malondialdehyde (MDA) and H2O2 content were determined by Lipid Peroxidation MDA Assay Kit and Hydrogen Peroxide Assay Kit (Beyotime Biotechnology Co., Ltd., Shanghai, China), respectively. The superoxide anion level was analyzed by Micro Superoxide Anion Assay Kit (Beijing Solarbio Science and Technology Co., Ltd., Beijing, China). All the experiments were performed according to the manufacturers’ protocols, and data were derived from three biological replicates. Student’s T-test was performed to calculate the p-values using SPSS 20.0 version.
Malondialdehyde (MDA) and H2O2 content were determined by Lipid Peroxidation MDA Assay Kit and Hydrogen Peroxide Assay Kit (Beyotime Biotechnology Co., Ltd., Shanghai, China), respectively. The superoxide anion level was analyzed by Micro Superoxide Anion Assay Kit (Beijing Solarbio Science and Technology Co., Ltd., Beijing, China). All the experiments were performed according to the manufacturers’ protocols, and data were derived from three biological replicates. Student’s T-test was performed to calculate the p-values using SPSS 20.0 version.
3
Superoxide and Antioxidant Enzyme Assays
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For superoxide staining, leaves were stained by nitroblue tetrazolium (NBT; 0.5 mg/ml) for 2 h and then soaked in boiling ethanol (95%) until the green color of the leaves faded. The Micro Superoxide Anion Assay Kit (BC1295, Solarbio, Beijing, China) was used to measure the superoxide content. The Catalase (CAT) Activity Assay Kit (BC0205, Solarbio), the Superoxide Dismutase (SOD) Activity Detection Kit (BC0170, Solarbio), the Peroxidase (POD) Activity Detection Kit (BC0090, Solarbio), and the Ascorbate Peroxidase (APX) Activity Assay Kit (BC0220, Solarbio) were used to detect the activity of CAT, SOD, POD, and APX, respectively.
4
Hydrogen Peroxide and Superoxide Anion Quantification
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The contents of H2O2 determination was calculated using a Hydrogen Peroxide Contents Detection Kit (Solarbio, Beijing, China) and O2− production determination was calculated using a Micro Superoxide Anion Assay Kit (Solarbio, Beijing, China) [47 (link)]. 3,30-Diaminobenzidine (DAB) was used to measure the levels of H2O2, as described in a previous study [48 (link)]. All experiments have been repeated independently three times with similar results.
5
Oxidative Stress Response in Maize Seedlings
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About 10-day-old maize seedlings were treated with 200 mM NaCl for 2 days, and the second leaves were harvested for the subsequent analysis. For H2O2 staining and O2− staining, the leaves were incubated in 1 mg ml−1 3,3′-diaminobenzidine (DAB) solution (pH 3.8, Coolaber, China) and 0.5 mg ml−1 nitroblue tetrazolium chloride (NBT) solution (Solarbio, China) for 8 h in dark at room temperature, respectively. After staining, the leaves were boiled in 95% ethanol for 10 min to decolorize, and then photographed. For quantification, the contents of H2O2 and O2− were determined using H2O2 Detection Kit (Leagene, China) and Micro Superoxide Anion Assay Kit (Solarbio, China) according to the manufacturer’s protocol, respectively.
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