Serum creatinine and BUN levels were determined using a biochemical autoanalyzer (Hitachi, Osaka, Japan). Renal MDA and 8-OHdG levels were analyzed using the MDA assay kit (Sigma-Aldrich, St. Louis, MO, USA) and the 8-OHdG ELISA kit (Abcam, Cambridge, MA, USA), respectively. Renal GSH and GSSG levels were analyzed using the GSH detection kit (Enzo Life Sciences, Farmingdale, NY, USA). Catalase and SOD activities were determined using colorimetric activity kits (Invitrogen, Carlsbad, CA, USA). Serum TNF-α and IL-6 levels were determined using ELISA kits (R&D Systems, Minneapolis, MN, USA). All assays were performed according to the manufacturers’ protocols.
Colorimetric activity kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Colorimetric activity kit is a laboratory instrument used to measure the optical density or absorbance of a sample. It provides a quantitative assessment of the concentration or activity of a specific analyte in a solution. The kit utilizes colorimetric detection methods to generate a measurable signal proportional to the target analyte's presence.
Automatically generated - may contain errors
Lab products found in correlation
Biochemical autoanalyzer, by Hitachi (2 mentions)
Gsh detection kit, by Enzo Life Sciences (2 mentions)
Mda assay kit, by Merck Group (2 mentions)
8 ohdg elisa kit, by Abcam (1 mentions)
8 ohdg assay kit, by Abcam (1 mentions)
Lipid peroxidation assay kit, by Merck Group (1 mentions)
Colorimetric detection kit, by Thermo Fisher Scientific (1 mentions)
5 protocols using colorimetric activity kit
1
Biomarkers of Renal Oxidative Stress
2
Oxidative Stress Biomarkers in Serum
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The serum concentrations of creatinine and BUN were measured using a biochemical autoanalyzer (Hitachi, Osaka, Japan). Catalase and SOD activities were analyzed using colorimetric activity kits (Invitrogen, Carlsbad, CA, USA). The serum concentrations of TNF-α and IL-6 were determined using ELISA kits (R&D Systems, Minneapolis, MN, USA). MDA and 8-OHdG levels were analyzed using an MDA assay kit (Sigma-Aldrich, St. Louis, MO, USA) and an 8-OHdG assay kit (Abcam, Cambridge, MA, USA), respectively. GSH levels were determined using a GSH detection kit (Enzo Life Sciences, Farmingdale, NY, USA). All analyses were carried out following the manufacturers’ protocols.
3
Measuring Protein Kinase A Activity in HEK293 Cells
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HEK293 cells were stimulated with thapsigargin (2 μm ), forskolin (50 μm ), or ionomycin (5 μm ) for 20 min. Cells were then lysed and protein kinase A activity was detected using a colorimetric activity kit (Invitrogen), following the supplier's instructions. Protein kinase A substrate was phosphorylated by the kinase in the presence of ATP. Absorbance was read at 450 nm using a microplate reader.
4
Quantitative Measurement of SOD Activity
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Total SOD activity was measured using an Invitrogen SOD colorimetric activity kit according to the manufacturer's instruction. This kit is designed to quantitatively measure all types of SOD activity, including Cu/Zn, Mn and FeSOD types according to the manufacturer's information. Briefly, 24 h after treatment, cells were washed with cold PBS twice, pelleted by centrifugation at 1,200 x g and room temperature for 5 min, homogenized and lysed using lysis buffer in the kit. The lysates were centrifuged at 1,200 x g at 4˚C for 10 min and the supernatants were used for SOD activity assessment. The optical density at 450 nm was read using a plate reader. All assays were performed in triplicate in three independent experiments.
5
Oxidative Stress Marker Analysis
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The following oxidative stress markers were assessed in this study: MDA, SOD, and GSH. MDA level was measured using lipid peroxidation assay kit (Sigma-Aldrich, Singapore; catalog number MAK085), SOD activity using colorimetric activity kit (Invitrogen, Carlsbad, CA; catalog number EIASODC), and GSH level with colorimetric detection kit (Invitrogen, Carlsbad, CA; catalog number EIAGSHC), all of which were performed to the manufacturers' instructions. The obtained concentration for each oxidative stress marker was normalized for tissue protein content, and MDA level and GSH and SOD concentrations were expressed as nmol/mg protein, μmol/mg protein, and U/mg protein, respectively.
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