The two new novel probes, Aminophenyl fluorescein (APF) and Hydroxyphenyl fluorescein (HPF) developed by Tetsuo Nagano et. al. (1), are selective for the detection of highly reactive oxygen species (hROS). Both probes have little reactivity towards other ROS such as: singlet oxygen (021), superoxide (O2-•), hydrogen peroxide (H202), nitric oxide (NO•), and alkyl peroxide (RO2•) (see table below)1. HPF/APF are cell permeable and can be used in combination to detect hypochlorite (-OCl) production in cells (see fig 1). Hypochlorite can be detected by loading two samples, one with APF and the other with HPF. Hypochlorite production is visualized by increase in fluorescence of APF loaded cells and no increase in fluorescence in HPF loaded cells.
- Quenched Cell permeable dye.
- One Step, No wash Homogenous assay.
- Adaptable to High throughput assay platforms.
- Can monitor multiple time points to follow real time kinetics.
- Non-destructive cell based assay allows monitoring of additional parameters.
- Applications – Fluorescent plate reader / Flow Cytometery / Fluorescent Microscopy.