Scientific multiskan go enzyme marker

HT-22 cells (1 × 10⁴ cells/well) were cultured in 96-well plates at 37 °C with 5% CO₂ and exposed to 500 μM H₂O₂ for 12 h. Cells treated with a culture medium were used as a control only. After removing the supernatant of each well and washing it twice with PBS, 20 μL of 5 mg/mL MTT reagent (Biovision Inc., Milpitas CA, USA) was introduced and incubated for 4 h. After the supernatant had been removed and 200 μL of DMSO had been added, the wells were well mixed. The absorbance intensity was measured at 570 nm with the Scientific Multiskan GO enzyme marker (Thermo, Shanghai, China). Relative cell viability was expressed as a percentage relative to untreated control cells.

A measurement of 100 mg of brain tissue was weighed, PBS was added, homogenized thoroughly, and centrifuged at 2000 r/min for 20 min, and the supernatant was collected. An ELISA kit (Jianglai Biologicals, Shanghai, China) was used for the assay according to the manufacturer’s instructions. The absorbance values of the samples were measured at 450 nm by a Scientific Multiskan GO enzyme marker (Thermo, Shanghai, China), and the concentration of each sample was obtained from the standard curve.