Assays for nitric oxide expression.
The purpose of this chapter is to review the analytical methodology specifically associated with studying the role of nitric oxide (NO) in mast cell physiology and biochemistry. The methodology for measuring cellular secretion of nitric oxide with Griess Reagent will be described in detail, as well as the use of 4,5-diaminofluorescein diacetate for continuous monitoring of nitric oxide production in live cells. We will point out the limitations of the analytical techniques and also indicate areas in which promising analytical techniques have not yet been applied to the study of mast cell physiology, that is, new research opportunities. In addition to reviewing the methodology associated with measuring NO itself, we will briefly touch upon some analytical methods important in characterizing the biochemical products formed from nitric oxide (e.g., 3-nitrotyrosine).
Stone
William L
WL
Department of Pediatrics, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
Yang
Hongsong
H
Qui
Min
M
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002478:Cells, Cultured; D003497:Cyclic N-Oxides / Q000378:metabolism; D005029:Ethylenediamines / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D016166:Free Radical Scavengers / Q000378:metabolism; D007093:Imidazoles / Q000378:metabolism; D008407:Mast Cells / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D015394:Molecular Structure; D009566:Nitrates / Q000032:analysis; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009573:Nitrites / Q000032:analysis; D013424:Sulfanilamides / Q000378:metabolism
D003497:Cyclic N-Oxides; D005029:Ethylenediamines; D005456:Fluorescent Dyes; D016166:Free Radical Scavengers; C095000:Griess reagent; D007093:Imidazoles; D009566:Nitrates; D009573:Nitrites; D013424:Sulfanilamides; C079391:2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase