Total malondialdehyde (MDA) levels in the liver were measured using a colorimetric microplate assay kit for thiobarbituric acid reactive substances (Oxford Biochemical Research, Oxford, MI, USA), as previously described [24 (link)]. Glutathione (GSH) levels in the liver were examined using a GSSG/GSH quantification kit (Dojindo Laboratories, Kumamoto, Japan), as previously described [22 (link)].
Gssg gsh quantification kit
Manufactured by Dojindo Laboratories
Sourced in Japan, United States, China
The GSSG/GSH Quantification Kit is a laboratory tool designed to measure the levels of glutathione disulfide (GSSG) and reduced glutathione (GSH) in samples. The kit provides a rapid and accurate method for quantifying these important cellular antioxidants. It is intended for use in research and scientific applications.
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Lab products found in correlation
Sod assay kit wst, by Dojindo Laboratories (10 mentions)
Oxiselect catalase activity assay kit, by Cell Biolabs (4 mentions)
Lactate assay kit wst, by Dojindo Laboratories (3 mentions)
5 sulfosalicylic acid dihydrate, by Fujifilm (3 mentions)
Mda assay kit, by Abcam (2 mentions)
5 sulfosalicylic acid solution, by Fujifilm (2 mentions)
Varioskan lux plate reader, by Thermo Fisher Scientific (2 mentions)
Glucose assay kit wst, by Dojindo Laboratories (2 mentions)
Cholesterol e test, by Fujifilm (2 mentions)
Nad nadh assay kit wst, by Dojindo Laboratories (2 mentions)
Tbars assay kit, by Cayman Chemical (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Multiskan go, by Thermo Fisher Scientific (2 mentions)
Rnalater, by Thermo Fisher Scientific (2 mentions)
Model 680 microplate reader, by Bio-Rad (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Tissuelyser system, by Qiagen (2 mentions)
Model 680, by Bio-Rad (1 mentions)
Nadp nadph assay kit, by Beyotime (1 mentions)
97 protocols using gssg gsh quantification kit
1
Oxidative Stress Biomarkers in Liver
2
Quantification of Glutathione Redox States
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An oxidized/reduced glutathione (GSSG/GSH) Quantification Kit (Dojindo, Kumamoto, Japan) was used according to the vendor’s protocol to quantify the amount of GSH [25 (link)]. Absorbance was measured using a microplate reader (Model 680, BioRad, Hercules, CA, USA).
3
Antioxidant Enzyme Activity in Macrophages
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Activity of antioxidant enzymes in macrophages was measured using the OxiSelect Catalase Activity Assay Kit (Cell Biolabs Inc, San Diego, CA) for catalase, GSSG/GSH Quantification kit (DOJINDO Inc. Rockville, MD) for total, oxidized and reduced GSH, and SOD Assay Kit-WST (DOJINDO Inc. Rockville, MD) to measure intracellular SOD activity according to the manufacturers' guidelines.
4
Antioxidant Activity Measurement in RBCs
5
Oxidative Stress Markers in Liver Tissue
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Liver tissue was excised from the mice at 7 d and 28 d after treatment. The levels of superoxide dismutase (SOD), total glutathione, DNA 8-hydroxy-2′-deoxyguanosine (8-OHdG), and malondialdehyde (MDA) in the liver tissue were measured as oxidative stress markers. SOD was measured using the SOD Assay kit–WST (Dojindo, Kumamoto, Japan), total glutathione was measured with a GSSG/GSH Quantification kit (Dojindo), 8-OHdG was measured with a High Sensitive 8-OHdG Check ELISA kit (JaICA, Shizuoka, Japan), and MDA was measured with a lipid peroxidation (MDA) assay kit (Abcam, Cambridge, England). All tests were conducted according to the manufacturer's instructions.
6
Quantifying Liver Glutathione Levels
7
Protein and Glutathione Quantification in Mucus
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Protein concentrations were determined using the BCA protein assay with BSA as a standard. The concentrations of CES-1 were determined using a CES-1 ELISA kit (RayBiotech; Peachtree Corners, GA, USA). Glutathione concentrations were determined using a GSSG/GSH quantification kit (DOJINDO, Kumamoto, Japan). Equal amounts of mucus collected from the left and right sides were mixed and analyzed except for one sample of INM mucus, as previously described.
8
Cardiac Glutathione Quantification
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The GSH and GSSG levels in cardiac tissues were measured using the GSSG/GSH quantification kit (Dojindo Molecular Technologies Inc., Kumamoto, Japan). Briefly, tissue pieces (50 mg) were homogenized with 1 mL of 5% (v/v) 5-sulfosalicylic acid, and the mixture was centrifuged at 8000× g for 10 min at 4 °C. Total GSH and oxidized GSH (GSSG) levels in the supernatant were determined according to the manufacturer’s protocol. Reduced GSH levels were calculated based on the concentrations of the total GSH and GSSG that were measured using the following formula: reduced GSH (µmol/L) = total GSH (µmol/L) − 2 × GSSG (µmol/L). The amount of GSH and GSSG was calculated using the standard curve.
9
Quantifying GSH and GSSG Levels
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GSH and GSSG concentrations were measured by using a GSSG/GSH Quantification Kit (Dojindo Molecular Technologies, Inc., Rockville, MD, United States). Jejunal segments were homogenized with 5% sodium sulfosalicylate (Wako) and centrifuged to remove proteins. For each sample, the supernatant was added to a well to which coenzyme working solution and enzyme working solution had been previously added, and incubated at room temperature. Then, the substrate working solution was added to the well and it was incubated at room temperature. The absorbance of samples and the GSH standard was measured at 405 nm using a microplate reader (TECAN GENios, Grodig, Austria). For measurement of GSSG, the supernatant was treated with masking solution and then added to a well as described above. The absorbance of samples and standard GSSG was measured at 405 nm.
10
RBC Glutathione Quantification Protocol
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RBC glutathione levels were measured using a GSSG/GSH Quantification Kit (Dojindo) according to the manufacturer’s protocol38 (link). Briefly, RBCs were hemolyzed with 10 times the amount of 5% 5-sulfosalicylic acid solution (FUJIFILM Wako), and the samples were centrifuged at 8000 × g for 10 min to remove proteins7 (link). The samples and buffer solution were mixed and incubated for 1 h at 37 °C. Then Substrate and Enzyme/coenzyme working solution was added. After 10 min of incubation, absorbance was measured at 412 nm using a Varioskan LUX plate reader (Thermo Fisher Scientific, Kanagawa, Japan). The number of mice in each group is shown in Supplementary Table S1 .
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