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包埋后免疫细胞化学技术揭示大鼠肝细胞中丝氨酸:丙酮酸氨基转移酶的精细定位

Fine localization of serine:pyruvate aminotransferase in rat hepatocytes revealed by a post-embedding immunocytochemical technique.

作者信息

Yokota S, Oda T

出版信息

Histochemistry. 1984;80(6):591-5.

PMID:6469715
Abstract

Immunocytochemical localization of serine:pyruvate aminotransferase (SPT) in rat hepatocytes was studied using a protein A-gold technique. Rat liver was fixed by perfusion. Vibratome sections (100 micron thick) of the liver were embedded in Epon or Lowicryl K4M. Ultrathin sections were incubated with antiSPT, followed by protein A-gold complex. Gold particles representing the antigenic sites for SPT were seen in three subcellular compartments, peroxisomes, mitochondria, and cytoplasm. In the control experiments the specificity of the immunolabelling was confirmed. Quantitative analysis of the labelling density showed that main subcellular compartments containing SPT are mitochondria and peroxisomes. In addition, the gold particles distributing in the cytoplasm were 16%-29% of the total labelling. The result indicated that the cytoplasm also contains SPT with a low density.

摘要

采用蛋白A-金技术研究了大鼠肝细胞中丝氨酸:丙酮酸氨基转移酶(SPT)的免疫细胞化学定位。大鼠肝脏通过灌注固定。将肝脏的振动切片(100微米厚)包埋在环氧树脂或Lowicryl K4M中。超薄切片用抗SPT孵育,然后用蛋白A-金复合物处理。在三个亚细胞区室,即过氧化物酶体、线粒体和细胞质中可见代表SPT抗原位点的金颗粒。在对照实验中证实了免疫标记的特异性。标记密度的定量分析表明,含有SPT的主要亚细胞区室是线粒体和过氧化物酶体。此外,分布在细胞质中的金颗粒占总标记的16%-29%。结果表明,细胞质中也含有低密度的SPT。

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