Description
This kit enables the detection of an early apoptosis event based on the measurement of the glutathione content in live mammalian cells using a fluorescence microplate reader. Diminished cellular glutathione (GSH), which is an important anti-oxidant in mammalian cells, occurs at the early stages of mitochondria-associated apoptosis because of GSH efflux. This kit uses monochlorobimane (MCB), a cell permeant dye that has a high affinity for GSH, to detect the cellular GSH level in an apoptotic event. The unreacted dye is almost non-fluorescent. After reaction of the dye with GSH, catalyzed by endogenous glutathione-S-transferase (GST) enzymes, the dye fluoresces blue, with excitation at 394 nm and emission at 490 nm. The intensity of the fluorescence signal generated from the assay reflects the amount of GSH that is present in the cells.