The level of superoxide anion (O2•−) and ROS were detected using a kit from Enzo life sciences (Tokyo, Japan, ENZ-51010) following the protocol provided by the manufacturer [32 (link),33 (link),35 (link)]. Briefly, cells in 96-well plates were preloaded with O2•− detection reagent (orange) and oxidative stress detection reagent (green) for 3 h, followed by stimulation with acrolein for 1 h. The fluorescent images of cells were captured using the immunofluorescent microscope (IX71, Olympus, Tokyo, Japan).
Enz 51010
Manufactured by Enzo Life Sciences
Sourced in Japan
The ENZ-51010 is a laboratory centrifuge designed for general-purpose applications. It features a compact and durable construction, with a maximum speed of 6,000 RPM and a maximum capacity of 4 x 100 mL. The centrifuge is equipped with an electronic speed control and a digital display for easy monitoring of the run parameters.
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Lab products found in correlation
5 protocols using enz 51010
1
Superoxide and ROS Detection Assay
2
Intracellular ROS Quantification by DCFHDA
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Intracellular ROS generation was measured using a DCFHDA fluorescent dye (Molecular Probes/Invitrogen, Thermo Fisher Scientific, Grand Island, NY, USA) and a Total ROS/Superoxide Detection Kit (ENZO life Sciences, Plymouth Meeting, PA, USA). The CPs cells were cultured in six-well plates at a density of 1 × 104/well. Following treatment with an appropriate concentration of CuE, the cells were incubated with 10 μM DCFH-DA at 37 °C for 30 min and then washed twice with PBS. ROS/superoxide concentration was assessed by staining ROS and superoxide detection reagent according to the protocol provided by the manufacturer (ENZ-51010, ENZO life Sciences). For each experiment, the cells were analyzed for fluorescence using flow cytometry. Data were analyzed using the WinMDI 2.8 free software.
3
Intracellular ROS Detection in Macrophages
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Intracellular ROS generation was measured using a total ROS/superoxide detection kit (Enzo Life Sciences Inc., Farmingdale, NY, USA). Macrophages (105 cells/well) were seeded in 96-well plates and were treated with LtxA (1 µg/mL) at 37 °C for 1 h in the absence or presence of cranberry PACs at different concentrations, as described above. ROS and superoxide generation was then assessed using a commercial kit (ENZ-51010, Enzo Life Sciences Inc.).
4
Superoxide Anion and ROS Detection
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The generation of superoxide anion (O2•－) and ROS was detected by using a commercially available kit from Enzo life sciences (Tokyo, Japan, ENZ-51010) following the manual of the manufacturer, as previously described in our previous reports [35 ,37 ]. Briefly, cells in 96-well plates were loaded with O2•－ detection reagent (orange) and oxidative stress detection reagent (green), followed by stimulation with DSS or H2O2 for 3 h. The fluorescent images were captured using the immunofluorescent microscope (IX71, Olympus, Tokyo, Japan).
5
Quantifying Superoxide and ROS Levels
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Superoxide anion and reactive oxygen species (ROS) levels were measured using a commercial kit (Cat #ENZ-51010, Enzo Life Sciences, Farmingdale, NY, USA) per the manufacturer’s protocol. Cells were seeded in 96-well plates and pre-loaded with the superoxide detection reagent (orange fluorescent) and oxidative stress detection reagent (green fluorescent) for 3 h. Cells were then stimulated with LPS or TNBS, either alone or in combination, for 1 h. Fluorescent images were captured using an inverted fluorescent microscope (IX71, Olympus Corporation, Tokyo, Japan) to visualize superoxide and ROS levels through orange and green fluorescence, respectively.
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