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[电子显微镜放射自显影中固定依赖性假象形成的体外研究]

[In vitro studies on the fixation dependent artifact formation in electron microscope autoradiography].

作者信息

Ockenfels H, Pilgrim C, Unsöld M

出版信息

Acta Histochem Suppl. 1981;24:301-7.

PMID:6165045
Abstract

The binding of 14C-labeled L-fucose to bovine serum albumin by glutaraldehyde was investigated. Separation of protein-bound radioactivity from free precursor was performed either by TCA precipitation, thin layer chromatography or gel chromatography on Sephadex columns. For a positive control, [14C] fucose was replaced by [14C] leucine which has repeatedly been reported to be subjected to artifactual retention in glutaraldehyde-fixed tissue. It is shown that, under conditions where substantial amounts of free leucine are bound to albumin, radioactivity from L-[14C]fucose is not detectable in the protein fractions.

摘要

研究了戊二醛介导的14C标记的L-岩藻糖与牛血清白蛋白的结合。通过三氯乙酸沉淀、薄层色谱或Sephadex柱凝胶色谱法将与蛋白质结合的放射性与游离前体分离。作为阳性对照,用[14C]亮氨酸替代[14C]岩藻糖,[14C]亮氨酸在戊二醛固定组织中反复被报道会出现人为滞留。结果表明,在大量游离亮氨酸与白蛋白结合的条件下,蛋白质组分中未检测到L-[14C]岩藻糖的放射性。

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