Ransod sd125

A method based on Hodges et al. [21 (link)] was used to determine malondialdehyde (MDA) content in brain tissue. A 100-μL brain sample or seven different concentrations (5,10, 20, 25, 50, 75 and 100 μM of the standard 1,1,3,3-tetramethoxypropane (TMP) were mixed separately with 200 μL 10% trichloroacetic acid (TCA), after which each mixture was reacted on ice for 15 min to precipitate protein, followed by centrifuging at 3200 g for 15 min (4 °C), collecting 200 μL supernatant, adding 200 μL of 0.67% thiobarbituric acid (TBA), reacting in a water bath (100 °C) for 10 min and measuring absorbance at 535 nm for MDA quantitation based on the standard curve.
The superoxide dismutase (SOD) activity was determined using a commercial kit (RANSOD-SD125, Randox Laboratories Ltd., Antrim, UK). The catalase activity was determined using a commercial kit (707002, Cayman Chemical Co, Ann Arbor, MI, USA). The glutathione content was determined using a commercial kit (703002, Cayman Chemical Co, Ann Arbor, MI, USA). The glutathione peroxidase (GSH-Px) activity was determined using a commercial kit (703102, Cayman Chemical Co, Ann Arbor, MI, USA). The acetylcholinesterase (AChE) activity was determined using a commercial kit (ab138871, Abcam Plc, MA, USA).

GPx and SOD activities were measured using ready-to-use assay kits in tissue homogenates (stomach, small intestine, liver, and kidney) (RANSEL RS505, RANSOD SD125, Randox Laboratories Ltd., Crumlin, County Antrim, UK) by using an automated BS-240 VET Clinical Chemistry Analyzer (Mindray, Shenzhen, China).
MDA concentrations were measured in tissue homogenates (stomach, small intestine, liver, and kidney) to assess lipid peroxidation status, using commercially available assay kits (TBARS Assay Kit, item no. 10009055, Cayman Chemicals, Ann Arbor, MI, USA). The principal measurement was based on the reaction with thiobarbituric acid (TBA) in boiling water for 60 min in an acidic medium, and measurement of the absorbance of the reaction mixture at 532 nm [28 (link)]. Absorbance was measured with VersaMax Tunable Microplate Reader (Molecular Devices, San Jose, CA, USA).