Monochlorobimane is widely used to determine GSH in living cells.

Store in dark at -20 ℃.

Protocol:

Stock solution preparation:

Prepare a stock solution of monochlorobimane at concentration of 40 mM in DMSO. Store at -20°C in dark.

1. Seed cells in 96-well plate and culture the cells overnight. You may treat the negative control well with cisplatin at 30 μM for 24h, which caused the characteristic glutathione depletion.

2. Prepare working solution of monochlorobimane at concentration of 40 μM in Dulbecco’s phosphate buffer (pH 7; 1 mM). Working solution should be prepared freshly before use.

3. Discard the culture medium. Add working solution in every well.

4. Incubate the cells at 37°C for 10 min in dark.

5. Measure the fluorescence at 394/490 nm. The glutathione levels were expressed as the percentage relative to glutathione levels in control cells (control = 100%).{{Čapek J, Hauschke M, Brůčková L, Roušar T. Comparison of glutathione levels measured using optimized monochlorobimane assay with those from ortho-phthalaldehyde assay in intact cells. J Pharmacol Toxicol M

Monochlorobimane is widely used to determine GSH in living cells. By adding monochlorobimane to cell culture medium and allowing intracellular glutathione S-transferases to form GSH–mCB adducts, GSH could be measured fluorometrically.

[1] Kamencic H, Lyon A, Paterson PG, Juurlink BH. Monochlorobimane fluorometric method to measure tissue glutathione. Anal Biochem. 2000 Nov 1;286(1):35-7. doi: 10.1006/abio.2000.4765. PMID: 11038270.