Overview

  • Product name

    Peroxynitrite Assay Kit (Cell-based)
    See all Peroxynitrite kits
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Due to its extremely short half-life and low steady-state concentration, it has been challenging to detect and understand the role of peroxynitrite (ONOO-) in biological systems. Peroxynitrite Assay Kit (Cell-based) (ab233468) has been developed to address this unmet need. It provides a sensitive tool to monitor ONOO- level in living cells. Peroxynitrite Sensor Green is developed as an excellent fluorescent probe, which can specifically react with intercellular ONOO- to generate a bright green fluorescent product. This kit can be used in fluorescence imaging and fluorescence microplate reader.

  • Notes

    Peroxynitrite is a strong oxidizing species and a highly active nitrating agent. Peroxynitrite is formed from the reaction between superoxide radicals and nitric oxide generated in cells. It can cause damages to a wide array of biomolecules including proteins, enzymes, lipids and nucleic acids, eventually contributing to cell death. Meanwhile, peroxynitrite can also have protective activities in vivo by contributing to host-defense responses against invading pathogens. Therefore, peroxynitrite is an essential biological oxidant involved in a board range of physiological and pathological processes.

  • Platform

    Microplate reader, Fluorescence microscope

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Assay Buffer 1 x 1ml
    DMSO 1 x 100µl
    Peroxynitrite Green reagent 1 vial

Images

  • RAW 264.7 cells at 100,000 cells/well/100 µL were seeded overnight in a Costar black wall/clear bottom 96-well plate. Cells were co-incubated with Peroxynitrite Sensor Green working solution and SIN-1 at the concentration from 50 to 200 µM at 37 ºC for 1 hour. Cells incubated with Peroxynitrite Sensor Green without SIN-1 treatment were used as control. The fluorescence signal were monitored at Ex/Em = 490/530 nm (cut off = 515 nm) with bottom read mode using a FlexStation microplate reader.

  • RAW264.7 cells at 100,000 cells/well/100 µL were seeded overnight in a Costar black wall/clear bottom 96-well plate. SIN-1 Treatment: Cells were co-incubated with Peroxynitrite Sensor Green working solution and 100 µM SIN-1 at 37ºC for 1 hour. Untreated control: The RAW 264.7 cells were incubated with Peroxynitrite Sensor Green without SIN-1 treatment. The fluorescence signals were measured using a fluorescence microscope with a FITC filter.

Protocols

References

ab233468 has not yet been referenced specifically in any publications.

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