Glutathione redox state (the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG); GSH/GSSG) was measured from 50 mg of the gastrocnemius muscle by GSH and GSSG commercial kits from Solarbio (BC1175 and BC1185, Beijing, China), according to the manufacturer’s protocols. Protein carbonyl content was assayed in the homogenate supernatant of 50 mg of gastrocnemius tissue using the commercial assay kit purchased from Solarbio (BC1275, Beijing, China), according to the manufacturer’s instructions.
Bc1185
Manufactured by Solarbio
Sourced in China
The BC1185 is a laboratory centrifuge designed for general use in research and clinical settings. It is capable of separating samples at high speeds to facilitate various laboratory processes.
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Lab products found in correlation
5 protocols using bc1185
1
Muscle Glutathione and Protein Carbonyl
2
Glutathione Redox State Measurement
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3
Quantifying Oxidative Stress in EA.hy926 Cells
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Following the LSS experiment, EA.hy926 cells were collected to measure secreted superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, glutathione (GSH) content and glutathione disulfide (GSSG) content using ELISAs (S8530, BC0025, BC1170, BC1185, Beijing Solarbio Science & Technology Co., Ltd.), according to the manufacturers’ protocols.
Following the LSS experiment, EA.hy926 cells were also incubated with the ROS-specific fluorescent dye dihydroethidium (DHE S0063, 50 μM, Beyotime Institute of Biotechnology). Following the incubation, the EA.hy926 cells were washed in PBS twice and then visualized and analyzed via fluorescence microscopy. The fluorescence intensity was semi-quantified from ≥3 random fields of view per slide from three different slides.
Following the LSS experiment, EA.hy926 cells were also incubated with the ROS-specific fluorescent dye dihydroethidium (DHE S0063, 50 μM, Beyotime Institute of Biotechnology). Following the incubation, the EA.hy926 cells were washed in PBS twice and then visualized and analyzed via fluorescence microscopy. The fluorescence intensity was semi-quantified from ≥3 random fields of view per slide from three different slides.
4
Antioxidant Activity Determination Protocol
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The centrifuge precipitate in Section 2.6 was dissolved with 1 mL 1 M NaOH containing 10% DMSO at 80 °C for 1 h were collected separately for antioxidant activity determination. The content of reduced glutathione (GSH), oxidized glutathione (GSSG), reactive oxygen species (ROS), and malondialdehyde (MDA), and the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) were measured using different assay kits that were all from Beijing Solarbio Science & Technology Co., Ltd., Beijing, China, and the item numbers were as follows: GSH by BC1175; GSSG by BC1185; ROS by CA1410; MDA by BC0025; SOD by BC0175; CAT by BC0205; GPX by BC1195.
5
Mitochondrial Function and Oxidative Stress Assays
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Superoxide dismutase activity assay Total superoxide dismutase (SOD) activity was tested by xanthine oxidase method with SOD Assay Kit (KeyGEN BioTECH, KGT001100, China) according to manufacturer's instruction.
Glutathione redox state Glutathione redox state was measured by total and oxidized glutathione (GSH and GSSG) respectively using a commercially available kit (Solarbio BC1170 and BC1185).
Mitochondria extraction and content determination Mitochondria of gastrocnemius were isolated with the Mitochondria Isolation Kit for Tissue (Invent, MM-038) according to the manufacturer's instruction and resuspended in D-PBS with protease inhibitors. Then mitochondria were quanti ed by BCA analysis.
Mitochondrial membrane potential determination Mitochondrial membrane potential was determined by investigating uorescence of 5,50,6,60-tetrachlore-1,10,3,30-tetraethylbenziml-dazolylcarbocyanine iodide (JC-1) (Solarbio, J8030) using a microplate uorometer at an excitation/emission wavelength of 485/590 nm.
Glutathione redox state Glutathione redox state was measured by total and oxidized glutathione (GSH and GSSG) respectively using a commercially available kit (Solarbio BC1170 and BC1185).
Mitochondria extraction and content determination Mitochondria of gastrocnemius were isolated with the Mitochondria Isolation Kit for Tissue (Invent, MM-038) according to the manufacturer's instruction and resuspended in D-PBS with protease inhibitors. Then mitochondria were quanti ed by BCA analysis.
Mitochondrial membrane potential determination Mitochondrial membrane potential was determined by investigating uorescence of 5,50,6,60-tetrachlore-1,10,3,30-tetraethylbenziml-dazolylcarbocyanine iodide (JC-1) (Solarbio, J8030) using a microplate uorometer at an excitation/emission wavelength of 485/590 nm.
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