• 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 - Fluorescence Plate Reader / Fluorescence Microscopy / Flow Cytometry

A new novel probe, Aminophenyl fluorescein (APF), developed by Tetsuo Nagano et. al. (1), is a general selective indcator for the detection of highly Reactive Oxygen Species (hROS). The probe has little reactivity towards other ROS such as: singlet oxygen (O21), superoxide (O2– •), hydrogen peroxide (H2O2), nitric oxide (NO•), and alkyl peroxide (RO2•) (see table below)1. APF is a cell permeable indicator that can be used to detect Hydroxyl Radical (.OH), Peroxynitrite: (ONOO- ) and hypochlorite (-OCl) production in cells.
Several fluorescent probes such as 2’,7’-dichlorodihydrofluorescein (DCFH) and di-hydrorhodamine123, have been developed to detect Reactive Oxygen Species (ROS). As Hempel and co-workers (2) have indicated, DCFH and di-hydrorhodamine123 can react with various ROS and oxidizing species such as hydrogen peroxide, hypochlorite, superoxide, nitric oxide, ferrous ion, and others. These probes should be considered as detecting a broad range of ROS (2).

Benzyl isothiocyanate-mediated generation of reactive oxygen species causes cell cycle arrest and induces apoptosis via activation of MAPK in human pancreatic cancer cells - Ravi P. Sahu, Ruifen Zhang, Sanjay Batra, Yan Shi and Sanjay K. Srivastava. Carcinogenesis (2009), Volume 30, Issue10, Pp. 1744-1753

Generation of highly-reactive oxygen species is closely related to hair cell damage in rat organ of corti treated with gentamicin - Y.H. Choung, A. Taura, K. Pak, S.J. Choi, M. Masuda and A.F. Ryan. Neuroscience, Volume 161, Issue 1, 16 June 2009, Pages 214-226

Post-translational activation of human phenylalanine 4-monooxygenase from an endobiotic to a xenobiotic enzyme by reactive oxygen and reactive nitrogen species - A. Antypa, C. Rebello, A. Biernacka, K. Krajewski, J. Cassam, S. C. Mitchell, G. B. Steventon. May 2010, Vol. 40, No. 5, Pages 319-330

Crocetin prevents retinal degeneration induced by oxidative and endoplasmic reticulum stresses via inhibition of caspase activity - Mika Yamauchi, Kazuhiro Tsuruma, Shunsuke Imai, Tomohiro Nakanishi, Naofumi Umigai, Masamitsu Shimazawa and Hideaki Hara. European Journal of Pharmacology, Volume 650, Issue 1, 10 January 2011, Pages 110-119

1. Ken-ichi Setsukinai, Yasuteru Urano, Katsuko Kakinuma, Hideyuki J. Majima , and Tetsuo Nagano. Development of Novel Fluorescence Probes That Can Reliably Detect Reactive Oxygen Species and Distinguish Specific Species. THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 278, No. 5, Issue of January 31, pp. 3170–3175, 2003
2. Hempel, S. L., Buettner, G. R., O’Malley, Y. Q., Wessels, D. A., and Flaherty, D. M. (1999) Free Radical Biol. Med. 27, 146–159