Intracellular gsh detection assay kit

Intracellular GSH level was assessed by an intracellular GSH detection assay kit (Abcam). Briefly, transfected cells were seeded in a 96-well plate at the density of 10⁴ per well overnight and treated with 100 µM FAC for 12 h before assessment. The intracellular GSH level was measured in accordance with the manufacturer’s instruction. Results were measured by flow cytometry (BD Biosciences).

The level of GSH was measured according to Intracellular GSH Detection Assay Kit (Abcam, USA). MDA-MB-231 and HCC1937 cells (1 × 10⁶) were treated with rimonabant, erastin, RSL3 as single treatment or their combination (rimonabant + erastin and rimonabant + RSL3) and then lysed in GSH assay buffer, and GSH levels were measured according to the manufacturer’s instructions. Colorimetric signals were measured by absorbance at 405 nm using a microplate reader (Molecular Devices, Sunnyvale, CA, USA). The GSH concentration was calculated using a standard curve. Protein quantification was performed using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, USA) as described in the manufacturer’s guidelines. The values were then normalized to total protein quantity per lysate sample. Results are expressed as means ± SD of fold changes relative to untreated controls.