This kit provides all necessary reagents, buffers and a detailed protocol for the detection of Reactive Oxygen Species (ROS, peroxidase activity) in live cells.

In healthy aerobic cells, ROS generation occurs at a controlled rate but under high stress conditions, its production is greatly increased, resulting in changes of many cell components including proteins and lipids.

Fluorogenic peroxidase substrates are converted to highly fluorescent products in the presence of the enzyme and hydrogen peroxide but can be relatively unstable for use in ELISAs. By forming the diacetate derivates, intracellular applications are more efficient, whereupon the acetates are cleaved by endogenous esterases, releasing the intact substrate. In the presence of nonspecific ROS, commonly produced during oxidative stress, the fluorescein substrate becomes oxidized, emitting green fluorescence.

The Live Cell Fluorescent Reactive Oxygen Species Detection Kit utilizes a cell permeable substrate that is a reliable fluorogenic marker for ROS detection. Upon enzyme activity, a highly fluorescent dye is produced, with EX: 495nm and EM: 530 nm. In addition, this kit also contains t-butyl hydroperoxide, a common ROS inducer, as a positive control.

The assay kit contains enough reagents to prepare up to 250 mL of staining solution, or approximately 100 assays (12-well tissue culture plate format), or 100 x 96-well tissue culture microtiterplate staining assays. Methods for testing various drug or inducer compounds are also described. This assay can be adapted for high-throughput, high-content assay screening analyses.

This kit is sold by Marker Gene under product code M1049.