Promega, V6911, GSH-Glo™ Glutathione Assay

GSH Depletion and Recovery in Treated Cells
HeLa cells (5000 cells/well in 96 well format) were exposed to l -Buthionine-sulfoximine (BSO). BSO inhibits GSH synthesis, thus reducing cellular GSH level.
BSO + Recovery: Cells treated with BSO for 20 hours, washed 2X with PBS and covered with fresh media without BSO. Cells assayed for GSH after 40 hours. 20hr BSO: Cells treated with BSO and assayed for GSH after 20 hours. 40hr BSO: Cells treated with BSO, washed at 20 hours and fresh media plus BSO added. Cells assayed for GSH after 40 hours.
The simple two-reagent-addition GSH detection assay minimizes the number of assay steps compared to conventional GSH assays and is easily adapted to higher throughput applications. No deproteination step is required, and results are achieved in as quickly as 30 minutes.
The luminescent method of the GSH-Glo™ Glutathione Assay avoids inherent background fluorescence associated with other methods, thereby providing excellent signal-to-background ratios. A half-life greater than 5 hours results in a stable signal, enabling batch processing of many plates at once using the GSH-Glo™ Assay.
The GSH-Glo™ Assay is recommended for measurement of total glutathione levels in samples. To measure the ratio, we recommend the GSH/GSSG-Glo™ Assay .