Peroxynitrite Detection Kit

1 Citation

The kit uses a novel probe (NP3) with excellent sensitivity and specificity to detect peroxynitrite in live cells and animals. The probe readily passes the blood-brain-barrier for neurological assessment.

Figure 1. (a) Fluorescent responses of NP3 (5 μM) toward various analytes (10 μM). Data shown represent fluorescent intensity at 470nm, 30 min after addition of the analytes. (b) ONOO− (final 10 μM) was quickly injected into a solution of NP3 (final 5μM), and the fluorescent intensity at 470 nm was plotted against time. (c) Fluorescence enhancement of NP3 (5μM) at 470 nm as a function of ONOO− (0−10 μM) after 15 min of reaction. All data acquired in PBS (10 mM, pH 7.4) with excitation at 375 nm.
Figure 2. (a) Time-lapse images taken from living EA.hy926 endothelial cells. Cells were preincubated with NP3 (5.0 μM), followed by stimulation with or without SIN-1 (0.5 mM). (b) Dynamic changes of NP3 fluorescence after SIN-1 (0.5 mM) treatment in panel a. (c) Effects of ONOO− scavengers uric acid (100 μM) and FeTTPS (1 μM) on changes in NP3 fluorescence in endothelial cells in the presence of ONOO− (60 μM). PI (red) stains nuclei. NP3 fluorescence was collected at 420−480 nm with λex 405 nm.
Figure 3. (a,b) Representative confocal images show time dependent accumulation of NP3 fluorescence (green; λex 405 nm, λem 420−480 nm) in (a) EA.hy926 cells following oxygen-glucose deprivation (OGD) exposure as well as in (b) human HBMEC cells. PI (red) stains nuclei. (c) EA.hy926 endothelial cells were pretreated with FeTTPS (1 μM), W7 (1 μM), melatonin (10 μM), or BAPTA (1 μM) 1 h prior to OGD treatment to suppress the ONOO− signal.
Figure 4. (a) Representative confocal images show temporal changes of ONOO−-dependent NP3 fluorescence (green; λex 405 nm, λem 420−480 nm) and MitoRed (red; λex 543 nm, λem 560−615 nm), as well as the apoptosis maker annexin V (blue; λex 488 nm, λem 505−550 nm), following OGD treatment. (b) Orthogonal projections onto the x−z (upper) and y−z (right) planes are shown to confirm the colocalization of NP3 and MitoRed throughout endothelial cells as shown in panel a. after ischemia injury.
Figure 5. Real-time visualization of endogenous peroxynitrite fluxes after brain microvessel injury with a combination of NP3 and in vivo two-photon laser scanning microscopy. (a, b) The images show dynamic NP3 fluorescence elevation (arrows) following (a) rose Bengal-induced vascular occlusion and (b) laser irradiation-induced vascular rapture in live mice. (c) Mean values of NP3 fluorescence intensity from panel b were used to quantify progressive ONOO− formation.
Figure 1. (a) Fluorescent responses of NP3 (5 μM) toward various analytes (10 μM). Data shown represent fluorescent intensity at 470nm, 30 min after addition of the analytes. (b) ONOO− (final 10 μM) was quickly injected into a solution of NP3 (final 5μM), and the fluorescent intensity at 470 nm was plotted against time. (c) Fluorescence enhancement of NP3 (5μM) at 470 nm as a function of ONOO− (0−10 μM) after 15 min of reaction. All data acquired in PBS (10 mM, pH 7.4) with excitation at 375 nm.
Figure 1. (a) Fluorescent responses of NP3 (5 μM) toward various analytes (10 μM). Data shown represent fluorescent intensity at 470nm, 30 min after addition of the analytes. (b) ONOO− (final 10 μM) was quickly injected into a solution of NP3 (final 5μM), and the fluorescent intensity at 470 nm was plotted against time. (c) Fluorescence enhancement of NP3 (5μM) at 470 nm as a function of ONOO− (0−10 μM) after 15 min of reaction. All data acquired in PBS (10 mM, pH 7.4) with excitation at 375 nm.
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Data Sheet
Protocol

SKU: FLHPF100-2

Size: 150 Tests
Price:
$298.00
Ships: 1-2 business days
  • Part# 4012: HPF, 1 Vial (2-8C)

    • Product Specific References

    PMID Publication
    31664127Zhu, Y, et al. 2019. Novel dihydroartemisinin dimer containing nitrogen atoms inhibits growth of endometrial cancer cells and may correlate with increasing intracellular peroxynitrite. Scientific Reports, 15528.

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