As per the manufacturer’s protocol, iron content kit, lipid peroxide product MDA kit and GSH kit (Abcam) were used to detect iron content, lipid peroxide MDA and GSH expression in tissues or cells, respectively.
Gsh kit
Manufactured by Abcam
Sourced in United States
The GSH kit is a laboratory tool used to detect and quantify the levels of glutathione (GSH), a crucial antioxidant found in cells. It provides a straightforward and reliable method for researchers to measure GSH concentrations in various biological samples.
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Lab products found in correlation
4 protocols using gsh kit
1
Tissue Iron, Lipid Peroxide, and GSH Analysis
2
Evaluating Iron and Oxidative Stress
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The levels of total iron (OD at 593 nm) and Fe2+ (OD at 593 nm) were measured in GC cells using an iron assay kit (ab83366, Abcam). MDA production (OD at 532 nm) was measured using (MDA) Kit (ab233471, Abcam). GSH levels (OD at 405 nm) were measured using GSH Kit (ab239709, Abcam). The above assays were performed using a FlexStation 3 multifunctional enzyme marker (Flexstation3, Molecular Devices, Sunnyvale, USA).
3
Glutathione Quantification by Colorimetry
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Glutathione levels were analyzed using GSH Kit from BioVision Inc. USA, using Colorimetric Detection according to the manufacturer’s instructions. The process is based on an enzymatic cycling method in the presence of GSH and a chromophore. The reduction of the chromophore produces a stable product, which can be followed kinetically at 450 nm. Therefore, its absorbance is directly proportional to the amount of GSH in the sample. The assay was performed using a spectrophotometer microplate reader, with 450 nm as the wavelength to measure the absorbance.
4
Quantification of Reduced Glutathione Levels
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Reduced GSH or glutathione levels were quantified using a commercially available GSH kit (BioVision, Mountain View, CA, United States of America) following the supplier’s instructions. Briefly, CAR-treated and untreated C33A cells were exposed to ice-cold GSH buffer (100 µl) for homogenization. The resulting homogenate was placed in a separate test tube containing chilled HClO₄ (10 ml) and vortexed for nearly 1 min. Subsequently, the homogenate was pelleted (13,000 × g for 2 min). The supernatant was collected and mixed with KOH in a ratio of 2:1, and after 5 min, the suspension was re-pelleted at the same force (13,000 × g), followed by the collection of supernatant for the remaining assay protocol. During the concluding steps, 10 ml of supernatant from different groups was diluted by reconstitution in 80 ml of assay buffer. Finally, the absorbance of fluorescence intensity was read at an excitation/emission ratio of 340/450 nm using a fluorimeter (Thermo-Fischer Scientific, United States of America).
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