Immunohistochemical Detection of S-Nitrosylated Proteins

  • Andrew J. Gow
  • Christiana W. Davis
  • David Munson
  • Harry Ischiropoulos
Part of the Methods in Molecular Biology™ book series (MIMB, volume 279)

Abstract

Accumulating evidence shows that S-nitrosothiols, formed by the addition of nitric oxide (NO) to a cysteine thiol, S-nitrosylation, are involved in basal cellular regulation. It has been proposed that SNO formation/removal may be disrupted in a variety of pathophysiological conditions. Two types of methodology are presently available to identify specific S-nitrosylated proteins: (1) derivatization and (2) post-purification chemical detection. Neither of these techniques allows for in situ visualization of SNOs. Recently, we demonstrated that an antibody generated to the SNO moiety could be used to detect SNO formation from each of three isoforms of NOS by immunohistochemistry. This chapter details the immunohistochemical methodology used to detect SNOs in situ, offering a potentially powerful alternative for detection of SNO within tissue sections.

Key Words

S-Nitrosothiols immunohistochemistry nitric oxide synthase nitrosylation 

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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Andrew J. Gow
    • 1
  • Christiana W. Davis
    • 2
  • David Munson
    • 2
  • Harry Ischiropoulos
    • 3
    • 4
  1. 1.Division of Neonatology, Joseph Stokes Junior Research Institute, and Abramson Research CenterChildren’s Hospital of PhiladelphiaPhiladelphia
  2. 2.Division of NeonatologyChildren’s Hospital of PhiladelphiaPhiladelphia
  3. 3.Division of Neonatology, Joseph Stokes Junior Research Institute, and Abramson Research CenterChildren’s Hospital of PhiladelphiaPhiladelphia
  4. 4.Department of Biochemistry and BiophysicsUniversity of Pennsylvania School of MedicinePhiladelphia

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