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Biotin Switch Assay Kit (S-Nitrosylation) (ab236207) uses a modification of the Jaffrey et al.

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Schematic Diagram - Biotin Switch Assay Kit (S-Nitrosylation) (AB236207), expandable thumbnail

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Key facts

Sample types

Tissue, Suspension cells, Adherent cells

What's included?

1 Kit
Components
S-Nitrosylation Blocking Reagent
3 x 1 Vial
S-Nitrosylation Buffer A
1 x 30 mL
S-Nitrosylation Buffer B (5X)
1 x 60 mL
S-Nitrosylation Detection Reagent I (HRP)
1 x 1 Vial
S-Nitrosylation Detection Reagent II (Fluorescein)
1 x 1 Vial
S-Nitrosylation Labeling Reagent
3 x 1 Vial
S-Nitrosylation Reducing Reagent
3 x 1 Vial
S-Nitrosylation Wash Buffer (10X)
1 x 25 mL

Recommended products

Biotin Switch Assay Kit (S-Nitrosylation) (ab236207) uses a modification of the Jaffrey et al.

Key facts

Sample types

Tissue, Suspension cells, Adherent cells

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

Biotin Switch Assay Kit (S-Nitrosylation) (ab236207) uses a modification of the Jaffrey et al. 'Biotin-switch' method to measure S-Nitrosylated (S-NO) proteins in whole cells or tissues by fluororescent microscopy, or in cell and tissue lysates by western blotting.

In the biotin switch assay method:
- firstly free SH groups are blocked (adding 125.1 amu per residue)
- then any S-NO bonds present as nitrosylated cysteines in the sample are cleaved
- the newly formed SH groups are then biotinylated (adding 523.6 amu per residue)
- the degree of biotinylation is then detected using either FITC-conjugated streptavidin for fluorometric detection, or HRP-conjugated streptavidin for colorimetric detection, and either western blotting or microscopy

Nitric oxide (NO) is produced by three distinct isoforms of nitric oxide synthase and functions as a key signaling molecule in physiology and pathophysiology. NO transduces its effects by reacting either directly with heme and non-heme centers of proteins or indirectly via further oxidation to various reactive nitrogen species (RNS).

Supplementary info

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

Activity summary

Nitric oxide (NO) is a small diatomic molecule with a mass of 30.01 Da. It plays a significant mechanical role as a signaling molecule in numerous systems. Nitric oxide is synthesized by nitric oxide synthases (NOS) and is expressed in endothelial cells neurons and various immune cells. Its production occurs in response to mechanical shear stress calcium influx or immunological stimulation. Nitric oxide exhibits high diffusibility enabling it to cross cellular membranes easily which is key for its function as a signaling messenger.

Biological function summary

Nitric oxide mediates vasodilation neurotransmission and immune response modulation. It is important for maintaining vascular homeostasis where it relaxes smooth muscle tissue promoting blood flow. In the central nervous system nitric oxide acts as a neurotransmitter and neuromodulator influencing synaptic plasticity. Within the immune system it functions to regulate leukocyte adhesion and can mediate microbial destruction. Nitric oxide often works within complexes associating with proteins like guanylate cyclase enhancing its signaling efficacy through nitrosylation.

Pathways

Nitric oxide integrates into key biological processes such as the nitric oxide signaling pathway and the endothelial nitric oxide pathway. It activates soluble guanylate cyclase increasing cyclic GMP levels leading to smooth muscle relaxation and vasodilation. In the endothelial pathway nitric oxide links with other proteins like endothelium-derived relaxing factor and interacts with oxidative stress pathways showing its role in cellular defense mechanisms. These pathways manage critical physiological responses and connect nitric oxide to other cardiovascular regulators.

Associated diseases and disorders

Nitric oxide dysregulation associates with conditions like cardiovascular disease and neurodegenerative disorders. A deficiency or excess production can contribute to hypertension or atherosclerosis given its role in vascular health. Neurodegenerative diseases such as Alzheimer's disease also show links to nitric oxide largely due to its involvement in neuroinflammation and synaptic dysfunctions. It interacts with proteins like amyloid-beta precursor protein and tau in these contexts highlighting its importance in pathology and therapeutic targets.

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1 product image

  • Schematic Diagram - Biotin Switch Assay Kit (S-Nitrosylation) (ab236207), expandable thumbnail

    Schematic Diagram - Biotin Switch Assay Kit (S-Nitrosylation) (ab236207)

    Derivitization of protein nitrosothiol (S-NO) with biotin.

    Protein free thiols are derivatized with a blocking agent. S-Nitrosothiols are reduced to yield free thiol(s) which are covalently labeled with maleimide-biotin. Subsequent detection by avidin-coupled reagents can be used to localize the biotinylated protein(s).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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