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NiSPY-3(Nitrative Stress Sensing Pyrromethene Dye)

NiSPY-3 (Nitrative Stress Sensing Pyrromethene Dye) is fluorescent reagent for detecting nitrosative stress. NiSPY-3 reacts with specifically peroxynitrite (ONOO-) when it works as detection reagent for reactive oxygen species.


  • NiSPY-3 reacts specifically with peroxynitrite (ONOO-) among other reactive oxygen species such as ・OH, O-・2 , H2O2, 1O2, NO etc. Fluorescent intensity is not increased by the existence of hydroxyl radical, singlet oxygen, hydrogen peroxide, hypochlorite, nitric oxide or superoxide.
  • Live cell fluorescent imaging is available with NiSPY-3.

Principle of the measurement

NiSPY-3 does not have fluorescence in neutral solution. When NiSPY-3 reacts with peroxynitrite, it becomes to have strong fluorescence (excitation: 490 nm, emission: 515 nm).


NiSPY-3  1mg
C23H19BF2N4O4   Mw:464.23

Preparation of the reagent
Dissolve all the reagent in 430 μL of DMSO (5mmol/L), and then dilute 500-5000 times with neutral buffer (final concentration: 1-10 μM) before use. We recommend to use up at once after probes are diluted.

NiSPY-3 (Nitrative Stress Sensing Pyrromethene Dye)

A)Fluorescence spectra of NiSPY-3 solution (10 μM NiSPY-3 in 0.1 M phosphate buffer pH 7.4  containing 0.1% DMF as a cosolvent) upon addition of peroxynitrite (final 0, 1, 2, 5,10, 20 μM).

B)Fluorescence response of NiSPY-3 in various ROS generation systems

R. Furuta, N. Kurake, K. Ishikawa, K. Takeda, H. Hashizume, H. Tanaka, H. Kondo, M. Sekine, M. Hori (2017)
Plasma Processes and Polymers, DOI: 10.1002/ppap.201700123
T. Ueno, Y. Urano, H. Kojima and T. Nagano: J. Am. Chem. Soc., 128, 10640-10641 (2006)
Code no. Product size Price Protocol MSDS
SK3003-01 NiSPY-3 1mg $586.00 Download