We assessed MDA (nmol/mL) by its reaction with 2-thiobarbituric acid (TBA) to produce a pink derivate quantifiable by ultra-high performance liquid chromatography with fluorescence detection (UHPLC-FD), using a method adapted from (Losdat et al. 2014 (link)). Details about the method can be found in the Supplementary Material . We assessed SOD activity (U/mL) using Cayman’s SOD assay kit (Cayman chemical company, USA), which is based on the detection of superoxide radicals generated by xanthine oxidase and neutralized by SOD. The reduced (GSH, ng/mL) and oxidized (glutathione disulfide GSSG, ng/mL) forms of glutathione were measured by ultra-high performance liquid chromatography tandem mass-spectrometry (UHPLC-MS/MS), according to Bouligand et al. (2006) with some modifications. Further details about the method can be found in the Supplementary Material . We quantified α, δ, γ-tocopherol in the plasma by UHPLC-MS/MS. More details about the method can be found in the Supplementary Material .
Cayman s sod assay kit
Manufactured by Cayman Chemical
Sourced in United States
Cayman's SOD assay kit is a laboratory tool designed to quantify superoxide dismutase (SOD) activity in a sample. It provides a reliable method for measuring the enzymatic activity of SOD, which plays a crucial role in the antioxidant defense system.
Automatically generated - may contain errors
5 protocols using cayman s sod assay kit
1
Oxidative Stress Biomarkers Quantification
2
Liver Antioxidant Assessment Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Approximately 100 mg of the liver samples were homogenized with 1 ml of designated solution for each analysis including total antioxidant capacity (TAC), glutathione (GSH) and oxidized GSH (GSSG), and superoxide dismutase (SOD) using a beads beater (Biospec Products, Bartlesville, OK, United States). The TAC of the liver tissues were determined using colorimetric microplate assay kits for Total antioxidant capacity (TA02, Oxford Biomedical Research, Oxford, MI, United States) according to Choi et al. (2020b) (link). The GSH and GSSG in the liver were analyzed by using Caymans GSH assay kits (Cayman Chemical, Ann Arbor, MI, United States) with a 20-time sample dilution. The activities of SOD in the liver were analyzed using Caymans SOD assay kit (Cayman Chemical) with a 400-time sample dilution. The supernatants of homogenized samples after the centrifugation at 4°C and 12,000 × g for 15 min were collected to analyze their protein content using Pierce BCA protein assay kits (Thermo Fisher Scientific) with a 20-time sample dilution. The TAC, GSH and GSSG concentrations, and SOD activities were expressed as values per mg protein.
3
Superoxide Dismutase Activity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cayman’s SOD Assay kit (Cayman Chemical Company, Ann Arbor, MI, USA, No. 706002), which uses tetrazolium salt for detection of superoxide radicals produced by xanthine oxidase and hypoxanthine, was used according to the manufacturer’s instructions [31 (link)]. This assay measures all forms of SOD (Cu/Zn, Mn, and FeSOD). A single unit of SOD is defined as the amount of the enzyme that is required in order to exhibit 50% dismutation of the superoxide radical. Plasma was diluted 1:5 with sample buffer before assaying for SOD activity. Results are expressed as units/mg of protein tested.
4
Superoxide Dismutase and Glutathione Peroxidase Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SOD activity was determined in order to detect superoxide radicals created from the xanthine/XO system by Cayman’s SOD assay kit (Cayman Chemical Co., Ann Arbor, MI, USA). The SOD activity was calculated and compared with the standard curve of SOD. The activity was expressed as units/mg protein. GPx activity was determined by the method of Hussain et al. (29 (link)). Briefly, supernatant from the liver tissue was incubated with the reaction mixture containing 48 mM sodium phosphate, 0.38 mM ethylenediaminetetraacetic acid, 0.12 mM β-nicotinamide adenine dinucleotide phosphate (NADP) hydrate, 0.95 mM sodium azide, 3.2 units of glutathione reductase, 1 mM glutathione, 0.02 mM DL-dithiothreitol, and 0.0007% H2O2. The measurement of the absorbance of NADP+ was performed at 340 nm compared to the reagent blank and standard curve of GPx. The values were expressed as units/mg protein. The protein concentration was measured following the method of Lowry et al. (30 (link)).
5
Measuring SOD Activity in Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As previously described [13 (link)] Cayman's SOD Assay kit (Cayman Chemical Company, Ann Arbor, MI, USA, No. 706002), which measures all forms of SOD activity (Cu/Zn, Mn and FeSOD), was used according to the manufacturer's instructions [34 (link)]. Plasma samples were diluted to 1:5 with Sample Buffer before for the assay. The results are reported as units/mg of protein tested.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!