Nitric Oxide Protocols

Carrying on the high standards of the much-praised first edition of Nitric Oxide Protocols, Aviv Hassid has brought together a panel of expert researchers and clinician scientists to describe in step-by-step detail the latest methodologies for the measurement of nitric oxide--and the enzyme that produces it-in biological tissues and fluids. The authors take advantage of the latest methodologies for the quantitation of biological fluids and tissues, including capillary electrophoresis, microcoaxial electrodes, in vivo measurement of nitric oxide in exhaled air, confocal microscopy, gas chromatography, in situ hybridization, and real-time polymerase chain reaction. Chapters on the measurement of the novel products of nitric oxide, such as nitrated proteins, S-nitrosylated proteins, and dioxygen-dependent NO metabolism, are also included. Additional chapters address the expression of nitric oxide synthase via the use of viral vectors in gene therapy for erectile dysfunction and cancer, as well as in retrovirus, adenovirus, or adenoassociated virus-mediated expression of nitric oxide synthase in vivo. The protocols follow the successful Methods in Molecular Biology(TM) series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
State-of-the-art and highly practical, Nitric Oxide Protocols, 2nd ed., offers investigators and clinican/scientists a gold-standard collection of readily reproducible analytical techniques for measuring levels of nitric oxide and determining its manifold functions and effects.

Includes supplementary material: sn.pub/extras

Zusammenfassung
A collection of techniques for measuring nitric oxide and the enzyme that produces it in biological tissues and fluids. It features methods that can be used to measure novel nitric oxide-related products such as protein nitration and nitrosation, as well as to express nitric oxide synthase in basic research and gene therapy using viral vectors.

Inhalt
Measurement of Nitric Oxide by Reconversion of Nitrate/Nitrite to NO.- High-Resolution Capillary Electrophoresis of Nitrite and Nitrate in Biological Samples.- Nitrite and Nitrate Measurements in Human Urine by Capillary Electrophoresis.- In Vivo Nitric Oxide Measurements Using a Microcoaxial Electrode.- Measurement of Exhaled Nitric Oxide.- Nitric Oxide Imaging in Neurons Using Confocal Microscopy.- Measurement of Nitric Oxide Synthase Activity In Vivo and In Vitro by Gas Chromatography-Mass Spectrometry.- A Nonradioactive Assay for Nitric Oxide Synthase Activity in Tissue Extracts.- Measurement of Nitric Oxide-Related Enzymes in the Brain by In Situ Hybridization.- Real-Time Polymerase Chain Reaction to Quantify mRNA for Endothelial Nitric Oxide Synthase.- Dioxygen-Dependent Metabolism of Nitric Oxide.- Proteomic Method for Identification of Tyrosine-Nitrated Proteins.- Immunohistochemical Detection of S-Nitrosylated Proteins.- Gene Therapy Techniques for the Delivery of Endothelial Nitric Oxide Synthase to the Corpora Cavernosa for Erectile Dysfunction.- Adenovirus-Mediated Nitric Oxide Synthase Gene Transfer Into the Nucleus Tractus Solitarius in Conscious Rats.- Gene Therapy in Cancer Via Use of a Retrovector Having a Tumor Specificity and Expressing Inducible Nitric Oxide Synthase.- Gene Transfer of Nitric Oxide Synthase Via the Use of Adeno-Associated Virus Vectors.- Adenovirus-Mediated Nitric Oxide Synthase Gene Transfer.