Invitrogen™ Premo™ Cellular Hydrogen Peroxide Sensor

The roGFP-Orp1 Chimera

Premo™ Cellular Hydrogen Peroxide Sensor combines the selectivity of a roGFP-Orp1 chimera with the transduction efficiency of BacMam 2.0 technology. Redox-sensitive green fluorescent protein (roGFP) detects redox potential through two cysteines introduced into the β-barrel structure of the GFP protein. Dilsulfide formation between the cysteines leads to protonation of GFP and increases the 400nm excitation spectra at the expense of the 488nm excitation spectra. The fluorescence at the two different excitation wavelengths can be quantitated using the same emission wavelength of 515nm. Orp1 is a hydrogen peroxide-sensitive yeast protein that forms disulfides upon reaction with hydrogen peroxide and has the ability to transduce signals to other redox proteins by disulfide exchange. By combining the hydrogen peroxide-selective properties of Orp1 and roGFP, it is possible to selectively quantitate hydrogen peroxide in cells.

BacMam Technology

With the incorporated BacMam technology, this sensor is ready to use—simply add to cells in growth media and incubate overnight. BacMam technology is based on the use of an insect cell virus (baculovirus) to efficiently deliver and express genes in mammalian cells. Because mammalian cells do not support replication of baculoviruses, transduction is extremely well tolerated and generally lacking in cytopathic effects, even at high virus levels.