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A sensitive and specific assay for superoxide anion released by neutrophils or macrophages based on bioluminescence of polynoidin.
Colepicolo, P; Camarero, V C; Nicolas, M T; Bassot, J M; Karnovsky, M L; Hastings, J W.
Afiliación
  • Colepicolo P; Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
Anal Biochem ; 184(2): 369-74, 1990 Feb 01.
Article en En | MEDLINE | ID: mdl-2158250
Using the luminescent protein polynoidin, present in the bioluminescent system isolated from the marine annelid Harmothoe lunulata, we have developed a new method to measure, specifically, superoxide anion (O2-) released by macrophages or neutrophils. A small quantity of an aqueous crude extract of polynoidin is used to detect O2- released by stimulated cells. Light emission is linearly dependent on the number of cells over a wide range (10(3) to 10(7) cells), and the assay is thus more sensitive than either luminol or ferricytochrome c reduction. Luminescence is enhanced 20% by mannitol, 80% by catalase, and is totally quenched by superoxide dismutase. For the same number of cells, neutrophils showed a threefold higher release of O2- and a twofold faster first-order light decay than stimulated macrophages, in accordance with data obtained by other methods.
Asunto(s)
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Superóxidos / Proteínas Luminiscentes / Macrófagos / Neutrófilos Tipo de estudio: Diagnostic_studies Límite: Animals Idioma: En Revista: Anal Biochem Año: 1990 Tipo del documento: Article Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Superóxidos / Proteínas Luminiscentes / Macrófagos / Neutrófilos Tipo de estudio: Diagnostic_studies Límite: Animals Idioma: En Revista: Anal Biochem Año: 1990 Tipo del documento: Article Pais de publicación: Estados Unidos