Assess changes in intracellular glutathione levels with the Intracellular GSH Assay. This whole cell glutathione assay employs a proprietary thiol-sensitive dye, ThioBright™ Green, to monitor relative changes in the concentration of the free form of glutathione, aka GSH. Analyze the fluorescent signal by flow cytometry.
- Prepare samples and controls.
- Reconstitute ThioBright™ Green with 500 µL DMSO to produce a 200X stock concentrate.
- Add ThioBright Green to each sample at 1:200. For example, add 5 µL to 995 µL cultured cells.
- Incubate 15-30 minutes.
- Optional: Spin and pellet cells, remove supernatants, and gently resuspend in Assay Buffer or other isotonic buffer.
- If desired, label with additional stains, such as Hoechst, Propidium Iodide, 7-AAD, or an antibody.
- Analyze with a flow cytometer. ThioBright Green excites at 490 nm and emits at 525 nm.