Titlebook: Nitric Oxide Protocols; Michael A. Titheradge Book 19981st edition Springer Science+Business Media New York 1998

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Cloning and Expression of Human eNOS and nNOS Using the Baculovirus-Insect Cell System,of NOS enzymes generate nitric oxide (NO) from arginine, producing citrulline as a co-product and requires the presence of heme, tetrahydrobiopterin (BH.), nicotinamide adenine dinucleotide phosphate (NADPH), calmodulin (CaM), flavin mononucleotide (FMN), and flavine adenine nucleotide (FAD) (.). Th

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Assay of NOS Activity by the Measurement of Conversion of Oxyhemoglobin to Methemoglobin by NO,itrulline, and NADP. (reviewed in ..). This chapter describes the measurement of NOS activity by utilizing the conversion of oxyhemoglobin to methemoglobin by NO while monitoring the absorption difference between the wavelengths 401 and 421 nm or at just 401 nm. This assay was first described by Fee

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Radiochemical Measurement of NOS Activity by Conversion of [,C],-Arginine to Citrulline Using HPLC ethod to assay for NOS activity. In cases where NOS is the sole or overwhelmingly predominant arginine-metabolizing enzyme present in a cell or tissue preparation, it is convenient to separate arginine from citrulline using a Dowex cation-exchange chromatography system (.., this volume). In some cel

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The Enzymatic Measurement of Nitrate and Nitrite,can be measured directly using NO-specific electrodes and chemiluminescence, the measurement of the stable end-products of NO metabolism, nitrite (NO..) and nitrate (NO..), is commonly used as a rapid and simple way to assess NO production when large numbers of samples are to be processed. NO can un

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Determination of NO with a Clark-Type Electrode,nderstanding of the complex biological functions of this widespread cellular messenger. Although several techniques are available for the sensitive detection of NO, most of these methods either involve sophisticated and expensive equipment (e.g., electron paramagnetic resonance [EPR] spectroscopy an

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The Measurement of NO in Biological Systems Using Chemiluminescence,scusses the chemiluminescence assay for NO and its intermediate and end products of oxidation—NO.. and NO... This is one of the most simple, sensitive, and accurate assays available. It will emphasize several practical aspects of the assay and give examples of recent applications, such as the study

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Measurement of eNOS and iNOS mRNA Expression Using Reverse Transcription Polymerase Chain Reaction,have been grouped into two broad categories: the constitutive Ca./calmodulin (CaM)-dependent endothelial NOS (eNOS) and neuronal NOS (nNOS) and an inducible Ca./CaM-independent NOS (iNOS). If NO is produced in insufficient amounts, or if it reacts with other compounds, it often escapes detection. Be