Hydroxyl / Peroxynitrite Detection Kit ™
Fluorescent Hydroxyl (•OH) / Peroxynitrite (ONOO -) Detection Kit

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.
Introduction:

A new novel probe, Hydroxyphenyl fluorescein (HPF), developed by Tetsuo Nagano et. al. (1), is a highly selective probe for the detection of highly Reactive Oxygen Species (hROS). It is a cell permeable highly sensitive fluorescent probe for hydroxyl radical (OH•), and peroxynitrite (ONOO-) detection. It has little reactivity towards other hROS such as: hypochlorite (-OCl), singlet oxygen (021), superoxide (02-•), hydrogen peroxide (H202), nitric oxide (NO•), and alkyl peroxide (RO2•) (see table below)1.

Assay Principle:


Reactivity:
Reactivity Profile of HPF:
ROS (RFU)

HPF (RFU)
Ex:499 Em:515

DCFH-DA (RFU)
Ex:500 Em:520
Hydroxyl Radical: OH
730 7400
Peroxynitrite: ONOO- 120
6600
Hypochlorite: -OCl 6 86
Oxygen Radical: 1O2 5 26
Superoxide: O2- 8 67
Hydrogen Peroxide : H202 2 190
Nitric Oxide: NO 6 150
Alkylperoxyl Radical: ROO 17 710
Autoxidation 1 2000
Citations

Paradoxical effect of l-arginine: Acceleration of endothelial cell senescence - Ortunato Scalera, Ellen I. Closs, Eva Flick, Jens Martens-Lobenhoffer, Jean P. Boissel, Uwe Lendeckel, Anke Heimburg and Stefanie M. Bode-Böger. Biochemical and Biophysical Research Communications, Volume 386, Issue 4, 4 September 2009, Pages 650-655


The neuroprotective effect of ladostigil against hydrogen peroxide-mediated cytotoxicity - Orly Weinreb, Orit Bar-Am, Tamar Amit, Noam Drigues, Yotam Sagi and Moussa B.H. Youdim. Chemico-Biological Interactions, Volume 175, Issues 1-3, 25 September 2008, Pages 318-326


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


The Novel Cholinesterase–Monoamine Oxidase Inhibitor and Antioxidant, Ladostigil, Confers Neuroprotection in Neuroblastoma Cells and Aged Rats - Orit Bar-Am, Orly Weinreb, Tamar Amit and Moussa B. H. Youdim. Journal of Molecular Neuroscience, Volume 37, Number 2, 135-145, DOI: 10.1007/s12031-008-9139-6


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 (doi:10.3109/00498251003675207)

References:
  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

Manufactured: by Daiichi Pure Chemicals Co. Ltd. Japan
Not for Sale in Japan

*Test size will depend on dilution and volume used per test. For example:

Dilution of HPF Volume of test Numer of tests
1:500 (10uM final) 0.2 mL 147
1: 1000 (5uM final) 0.2mL 294

 ProductCatalog #ContentsSize*Price
Hydroxyl / Peroxynitrite Detection KitFLHPF 100-2HPF 1 vial150$245.00

* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing