Michael Titheradge provides both the experienced researcher and the newcomer with time-tested techniques to cover all aspects of NO synthase and metabolism. Topics range from the cloning and expression of the different isoforms of NO synthase to the role of NO in DNA damage and apoptosis and include indirect and direct measurements of NO production, measurement on NO synthase mRNA both in situ and in vitro, and Western blotting and immunohistochemical localization of different isoforms of NO synthase. A variety of alternative techniques are described to enable researchers to set up an assay using either the simplest equipment or the most expensive EPR machines.Although the complexity of the samples leads to many spurious peaks (which often obscure peaks attributable to arginine and/or its metabolites), ... If the reaction-stopping protocol recommended here is used (see Subheading 3.1., item 6), sample pH will not be a problem. If samples for HPLC analysis have been otherwise treated, add TFA to 5% v/v (centrifuge to pellet protein if necessary) and proceed.
|Title||:||Nitric Oxide Protocols|
|Author||:||Michael A Titheradge|
|Publisher||:||Springer Science & Business Media - 1997|