The production of reactive oxygen species (ROS) was assayed with an ROS detection reagent kit (Nanjing Jiancheng Bioengineering Research Institute, China) according to the manufacturer's instructions. Chondrocytes were treated with JEZTC or GA for 1h followed by IL-1β-stimulation for 24h, then washed twice with PBS and incubated with dichlorodihydrofluorescein diacetate (DCFDA, 10 μM) in PBS in the dark for 60 min. After washing with PBS, the fluorescence was measured immediately using a fluorescence microplate reader (FLx800, Biotek, USA) with excitation and emission at 485 and 525 nm, respectively. At 4 and 8 weeks post-treatment, cartilage homogenate was used to detect ROS generation under frozen conditions. After centrifugation, the supernatant was incubated with DCFDA, and the experiments were performed as previously described.
Ros detection reagent kit
Manufactured by Nanjing Jiancheng
Sourced in China
The ROS detection reagent kit is a laboratory equipment product designed to detect the presence of reactive oxygen species (ROS) in various samples. The kit includes the necessary reagents and protocols to perform ROS detection experiments. The core function of this product is to provide researchers with the tools to quantify and analyze ROS levels in their samples.
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Lab products found in correlation
2 protocols using ros detection reagent kit
1
Quantification of Chondrocyte ROS Production
2
Intracellular ROS Production Assay
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Production of intracellular ROS was assayed with a ROS detection reagent kit (Nanjing Jiancheng Bioengineering Research Institute, Nanjing, China) according to the manufacturer's instructions. Briefly, primary osteoblasts were seeded in concentrations of 2×104 cells/well in 6-well plates and cultured with different concentrations of ZXHA-TC after 24 hours. All samples were done in triplicate. After 2, 4, and 6 days of treatment, culture medium was replaced with 10 µM 2,7-dichlorofuorescin diacetate and incubated for 30 min at 37℃ in the dark environment. After being washed with PBS three times, cells were detached using 0.25% trypsin-EDTA and suspended in a culture medium at a density of 1×105 cells/mL. Samples at 100 µL were randomly extracted from each of three parallel wells at the same drug concentration three times and transferred to 96-well plates. Oxidative burst was detected with a microplate fluorescence reader (FLx800, Biotek, Montpelier, VT, USA) with excitation and emission setting at 485 nm and 525 nm respectively.
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