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AB233468

Peroxynitrite Assay Kit (Cell-based)

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(5 Publications)

Peroxynitrite Assay Kit ab233468 uses the fluorescent Peroxynitrite Green dye (Ex/Em 490/530 nm) to assay peroxynitrite. Readout is by fluorometric plate reader or microscope.
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Functional Studies - Peroxynitrite Assay Kit (Cell-based) (AB233468)
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Supplier Data

Functional Studies - Peroxynitrite Assay Kit (Cell-based) (AB233468)

Detection of peroxynitrite in living cells upon SIN-1 treatment using ab233468 Peroxynitrite Assay Kit (Cell-based)

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 °C 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.

Immunocytochemistry/ Immunofluorescence - Peroxynitrite Assay Kit (Cell-based) (AB233468)
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Supplier Data

Immunocytochemistry/ Immunofluorescence - Peroxynitrite Assay Kit (Cell-based) (AB233468)

Fluorescence images of intracellular peroxynitrite in RAW 264.7 macrophage cells using ab233468 Peroxynitrite Assay Kit (Cell-based)

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 37°C 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.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based

Assay Platform

Microplate reader, Fluorescence microscope

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Product details

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.

Other 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.

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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Peroxynitrite is a reactive nitrogen species with a significant role in cellular signaling. It forms rapidly through the reaction of nitric oxide (NO) and superoxide (O2-) making it a critical part of oxidative stress mechanisms. Peroxynitrite is not a protein but a small molecule so it doesn't have a mass like proteins do. Despite being a molecule it has a significant presence in different cell types especially those under oxidative stress conditions. In interactions it often measures cellular responses to changes in redox balance.
Biological function summary

Peroxynitrite exerts its influence through oxidizing and nitrating various biological macromolecules. It primarily targets proteins lipids and DNA affecting their function and integrity. Although not part of a complex itself it disrupts cellular complexes like mitochondrial electron transport chain components. This prevention of proper ATP production highlights peroxynitrite's role in energy metabolism disruptions.

Pathways

Peroxynitrite plays a role in inflammatory and apoptotic pathways. It interacts with signaling molecules like NF-kB and caspases mediating cellular stress responses. Peroxynitrite modifies tyrosine residues on proteins leading to altered signaling functions. It closely relates to proteins such as SOD (superoxide dismutase) and iNOS (inducible nitric oxide synthase) due to its formation and effects on cellular processes.

Peroxynitrite relates to conditions like neurodegenerative diseases and cardiovascular diseases. Elevated peroxynitrite levels in these diseases indicate oxidative stress and damage to cellular components. During disorders like Alzheimer's disease it interacts with proteins such as amyloid-beta promoting neurotoxic effects. In cardiovascular diseases peroxynitrite interacts with proteins involved in endothelial dysfunction exacerbating disease processes. The presence of antioxidants and scavengers to mitigate peroxynitrite's effects reveal its importance in disease management strategies.
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Product protocols

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Target data

See full target information Oxido nitrite
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Publications (5)

Recent publications for all applications. Explore the full list and refine your search

Journal of diabetes and its complications 38:108832 PubMed39116474

2024

Methylglyoxal induces death in human brain neuronal cells (SH-SY5Y), prevented by metformin and dapagliflozin.

Applications

Unspecified application

Species

Unspecified reactive species

Samantha Victor-Sami,Ali Kamali-Roosta,Yousif A Shamsaldeen

Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics 21:e00301 PubMed38241160

2024

Nicotinamide riboside, pterostilbene and ibudilast protect motor neurons and extend survival in ALS mice.

Applications

Unspecified application

Species

Unspecified reactive species

Rafael López-Blanch,Rosario Salvador-Palmer,María Oriol-Caballo,Paz Moreno-Murciano,Ryan W Dellinger,José M Estrela,Elena Obrador

Redox biology 67:102929 PubMed37856999

2023

Arginase2 mediates contrast-induced acute kidney injury via facilitating nitrosative stress in tubular cells.

Applications

Unspecified application

Species

Unspecified reactive species

Ling-Yun Zhou,Kun Liu,Wen-Jun Yin,Yue-Liang Xie,Jiang-Lin Wang,Shan-Ru Zuo,Zhi-Yao Tang,Yi-Feng Wu,Xiao-Cong Zuo

Scientific reports 13:14223 PubMed37648766

2023

Impact of weak radiofrequency and static magnetic fields on key signaling molecules, intracellular pH, membrane potential, and cell growth in HT-1080 fibrosarcoma cells.

Applications

Unspecified application

Species

Unspecified reactive species

Hakki Gurhan,Frank Barnes

Redox biology 43:101989 PubMed33940548

2021

Lysosomal nitric oxide determines transition from autophagy to ferroptosis after exposure to plasma-activated Ringer's lactate.

Applications

Unspecified application

Species

Unspecified reactive species

Li Jiang,Hao Zheng,Qinying Lyu,Shotaro Hayashi,Kotaro Sato,Yoshitaka Sekido,Kae Nakamura,Hiromasa Tanaka,Kenji Ishikawa,Hiroaki Kajiyama,Masaaki Mizuno,Masaru Hori,Shinya Toyokuni
View all publications
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