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离体牛肾上腺嗜铬细胞儿茶酚胺分泌的直接荧光测定法

Direct fluorometric assay of catecholamine secretion from isolated bovine adrenal chromaffin cells.

作者信息

Lelkes P I, Friedman J E, Rosenheck K

出版信息

J Neurosci Methods. 1985 May;13(3-4):249-55. doi: 10.1016/0165-0270(85)90073-1.

Abstract

The intrinsic fluorescence of catecholamines can be exploited for a simple and speedy fluorescence assay of secretion from bovine adrenal chromaffin cells. Catecholamines are separated from concomitantly secreted proteins by centrifugation in 0.4 M perchloric acid, and their fluorescence is measured directly in a conventional spectrofluorometer without chemical derivatization. Catecholamine release, assayed by this technique, is virtually identical to data obtained in parallel by the classical trihydroxyindole method. Intrinsic catecholamine fluorescence does not differentiate between epinephrine and norepinephrine. Interferences from small molecular weight compounds not precipitated in perchloric acid are rare, and can be accounted for using appropriate calibration curves.

摘要

儿茶酚胺的固有荧光可用于对牛肾上腺嗜铬细胞分泌进行简单快速的荧光测定。通过在0.4M高氯酸中离心,可将儿茶酚胺与同时分泌的蛋白质分离,并且无需化学衍生化,直接在传统荧光分光光度计中测量其荧光。用该技术测定的儿茶酚胺释放与通过经典三羟基吲哚法并行获得的数据几乎相同。儿茶酚胺的固有荧光无法区分肾上腺素和去甲肾上腺素。未在高氯酸中沉淀的小分子化合物产生的干扰很少,可使用适当的校准曲线进行解释。

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