Fluoro hROS





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).

Key Benefits

  • 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.

Additional information

Kit Size




Readable Documents

Title Name
Protocol hROS Protocol.pdf
Datasheet hROS Datasheet.pdf
msds msds hROS.pdf

Kit contents and storage

Part# Reagent Temperature
Part# 4011 APF Dye 2-8°C


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
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



Title File Link Author(s) Journal Year; Edition:Pages
Benzyl isothiocyanate-mediated generation of reactive oxygen species causes cell cycle arrest and induces apoptosis via activation of MAPK in human pancreatic cancer cells http://carcin.oxfordjournals.org/content/30/10/1744.short 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 http://www.sciencedirect.com/science/article/pii/S0306452209003352 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 http://informahealthcare.com/doi/abs/10.3109/00498251003675207 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 http://www.sciencedirect.com/science/article/pii/S0014299910010071 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