Overview

  • Product name

    Biotin Switch Assay Kit (S-Nitrosylation)
    See all Nitric oxide kits
  • Sample type

    Tissue, Adherent cells, Suspension cells
  • Product overview

    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


     

  • Notes

    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).

     

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 kit
    S-Nitrosylation Blocking Reagent 3 vials
    S-Nitrosylation Buffer A 1 x 30ml
    S-Nitrosylation Buffer B (5X) 1 x 60ml
    S-Nitrosylation Detection Reagent I (HRP) 1 vial
    S-Nitrosylation Detection Reagent II (Fluorescein) 1 vial
    S-Nitrosylation Labeling Reagent 3 vials
    S-Nitrosylation Reducing Reagent 3 vials
    S-Nitrosylation Wash Buffer (10X) 1 x 25ml
  • Research areas

  • Relevance

    Nitric oxide (NO) is a key vertebrate biological messenger, playing an important role in neurotransmission, vascular regulation, immune responses and apoptosis. NO , also known as "endothelium-derived relaxing factor" or "EDRF", is synthesized from L-arginine, oxygen and NADPH by various NO synthases. Most of the NO in the cell is oxidized to nitrite (NO2-) and nitrate (NO3-), and therefore the concentrations of these anions are generally as a quantitative measure of NO production.

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Protocols

References

ab236207 has not yet been referenced specifically in any publications.

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