BCPAP cells or C643 cells were treated with or without L-BSO (50 μM; MedChemExpress, cat #HY-106376A) for 48 h before collection. GSH detection was performed according to the manufacturer's instructions (Beyotime, cat #S0053). Briefly, cells were washed with 1× PBS and collected, resuspended in 3 times the volume of protein removal reagent M solution. Cell samples were treated with two rapid freeze–thaw cycles using liquid nitrogen and 37°C water bath. The corresponding detection reagents were added to an appropriate amount of the cell sample. After 25 min, GSH was detected using a microplate analyzer at an absorbance of 412 nm. The GSH content was calculated according to the standard curve.
S0053
Manufactured by Beyotime
Sourced in China, United States
The S0053 is a laboratory instrument designed for performing electrochemical experiments. It is a potentiostat/galvanostat that can be used to control and monitor the potential and current of an electrochemical cell. The device is capable of various electrochemical techniques, such as cyclic voltammetry, chronoamperometry, and electrochemical impedance spectroscopy.
Automatically generated - may contain errors
Lab products found in correlation
S0131, by Beyotime (8 mentions)
Glutathione (gsh), by Beyotime (3 mentions)
S0101, by Beyotime (3 mentions)
A039 2 1, by Nanjing Jiancheng (2 mentions)
Ripa lysis buffer, by Beyotime (2 mentions)
Du800, by Beckman Coulter (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Multiskan go, by Thermo Fisher Scientific (2 mentions)
Microplate analyzer, by Bio-Rad (1 mentions)
A006 2 1, by Nanjing Jiancheng (1 mentions)
Inverted fluorescent microscope, by Olympus (1 mentions)
Bc3595, by Solarbio (1 mentions)
Kit s0109, by Beyotime (1 mentions)
Bc0205, by Solarbio (1 mentions)
Leica microtome, by Leica camera (1 mentions)
A003 4 1, by Nanjing Jiancheng (1 mentions)
S0021, by Beyotime (1 mentions)
Iron detection kit, by Nanjing Jiancheng (1 mentions)
E bc k139 s, by Elabscience (1 mentions)
Pi523, by Beyotime (1 mentions)
48 protocols using s0053
1
Quantifying Glutathione in Thyroid Cells
2
Ferroptosis-induced Lipid Peroxidation Assay
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Cellular lipid peroxide levels were assessed by BODIPY-C11 immunostaining. After living HK2 cells were seeded in laser confocal petri dishes, the cells were treated with erastin and/or vitexin. The dishes were removed from the incubator, and the cells were stained with the BODIPY-C11 581/591 sensor (D3861, Thermo Fisher, MA, USA) according to the manufacturer’s instructions. Different fluorescence emission peaks were observed by a Carl Zeiss confocal microscope. The levels of malondialdehyde (MDA), glutathione (GSH), and glutathione disulfide (GSSG) correlated with membrane lipid peroxidation during ferroptosis. Renal tissues or HK2 cells were homogenized in extraction buffer. MDA activity was detected with a customized commercial kit (S0131, Beyotime, Shanghai, China). The relative ratio of GSH/GSSG was assessed by a quantitative kit (S0053, Beyotime, Shanghai, China). The absorbances of MDA and GSH/GSSG were measured with an automated Bio-Rad microplate analyzer using a 96-well plate.
3
Oxidative Stress Markers in Neuronal Cell Lines
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The GSH and MDA content was measured in WT-SY5Y, Vector-SY5Y, and FtMt-SY5Y cell lines after OGD/R treatment using commercial kits (#S0053, #S0131M, Beyotime, Shanghai, China). The evaluation was carried out according to the protocol provided by the manufacturer. For GSH assessment, cells were washed with PBS and then centrifuged to collect the cells. First, the protein removal reagent solution was added to the cells and two rapid freeze-thaw cycles were performed on the samples. Afterwards, the samples were centrifuged at 10000×g for 10 min. The supernatant was mixed with the GSH assay regent and incubated at 25 °C for 5 min. Finally, the absorbance at 412 nm was measured using a microplate reader. The GSH content of each group was calculated based on a the standard curve. The MDA content was assessed using the thiobarbituric acid (TBA) method, which is based on spectrophotometric measurement of the product of the reaction of TBA with MDA. The MDA concentrations were calculated by the absorbance of the product at 532 nm. For detailed instructions, see supplementary document 1.
4
Intracellular ROS and GSH/GSSG Assay
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EJ and 5637 cells were cultured in 6-well plates overnight. Then, using fresh medium containing the indicated concentration of MTX-211, we replaced the growth medium and cultured for 48 h. An ROS assay kit (S0033S, Beyotime, Shanghai, China) was used to detect the intracellular ROS levels, and the protocol was as follows: The treated cells were incubated with 1.5 mL DCFH-DA (5 μM) for 20 min in 37 °C. Then, the serum-free medium was used to wash the cells three times, and the images were acquired using a fluorescent inverted microscope (Olympus, Tokyo, Japan). The levels of GSH and GSSG (oxidized GSH) in 5637 and EJ cells were determined using a GSH and GSSG assay kit (A006-2-1, Nanjing Jiancheng Bioengineering Institute, China, and S0053, Beyotime, Shanghai, China), respectively, as per the manufacturer’s instruction.
5
Liver Oxidative Stress Measurement
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The levels of malondialdehyde (MDA) and glutathione (GSH) were used to measure the degree of liver oxidative stress. Lipid peroxidation product level of lipid peroxides (MDA, S0131S, Beyotime, China) were estimated by using TBARS assay. The reduced glutathione (GSH, S0053, Beyotime, China) level was measured by according to the method of study (Beutler et al., 1963 (link)).
6
Quantification of Antioxidant Markers in HT-29 Cells
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HT-29 cells were seeded in 6-well plates and treated with different stimuli. Then, the collected cells were disrupted with ultrasound and centrifuged at 14,000 rpm for 15 min at 4°C. The supernatant was used for the colorimetric determination of SOD activity (S0101S, Beyotime Biotechnology, China) and H2O2 (BC3595, Solarbio, China) and GSH (S0053, Beyotime Biotechnology, China) contents. The experimental operation was carried out according to the manufacturer’s instructions.
7
Antioxidant Enzyme Activity Assay
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HeLa cells were collected and washed twice with PBS before lysis. The lysates were ultracentrifuged at 12,000 g for 30 min. The activities of antioxidant enzymes (SOD and GSH) were measured according to the manufacturer’s protocol (Kit S0109 and S0053, Beyotime Institute of Biotechnology, PR China).
8
Glutathione Redox Assay in HeLa Cells
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GSH/GSSG assay was measured with a kit (Beyotime, S0053). HeLa cells were treated with DMSO, Cu, HYF127c, HYF127c/Cu, and H2O2 for 12 h at 37 °C in an atmosphere of 5% CO2.
9
Cardiac Oxidative Stress Analysis
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After recanalization, the rat heart left ventricle anterior wall was cut. MDA, iron levels (A039-2-1, A039-2-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China), and the GSH/GSSG ratio (S0053, Beyotime, Shanghai, China) were assessed using their respective assay kits. The concentration of protein was detected by a BCA kit and the absorbance was determined with a microplate reader.
10
GSH Quantification Protocol
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GSH level was determined with a commercial kit (S0053, Beyotime Biotechnology, China) via procedures described in the manual.
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