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脑网格蛋白:培养细胞和组织中的免疫荧光模式

Brain clathrin: immunofluorescent patterns in cultured cells and tissues.

作者信息

Bloom W S, Fields K L, Yen S H, Haver K, Schook W, Puszkin S

出版信息

Proc Natl Acad Sci U S A. 1980 Sep;77(9):5520-4. doi: 10.1073/pnas.77.9.5520.

Abstract

High-purity bovine brain clathrin elicited antibodies, which were separated by affinity chromatography on clathrin conjugated with CNBr-activated Sepharose 4B. Indirect immunofluorescent labeling on cultured rat fibroblasts showed the characteristic dotted fluorescence throughout the entire cytoplasm and concentrated patches of fluorescence in areas where the Golgi apparatus and GERL (Golgi apparatus-endoplasmic reticulum-lysosome complex) exist next to the nucleus. Cerebellar tissue sections exhibited profuse immunofluorescence in the granular layer; dissociated cultures of neonatal rat cerebellar cells showed intense fluorescence in the cytoplasm and processes of granule cell neurons. The data suggest that clathrin is an abundant protein distinctly localized in the cytoplasm of cells and clearly correlated in its distribution and arrangement with actin stress fibers.

摘要

高纯度牛脑网格蛋白引发了抗体,这些抗体通过在与溴化氰活化的琼脂糖凝胶4B偶联的网格蛋白上进行亲和层析而分离。对培养的大鼠成纤维细胞进行间接免疫荧光标记显示,在整个细胞质中呈现出特征性的点状荧光,并且在细胞核旁存在高尔基体和GERL(高尔基体-内质网-溶酶体复合体)的区域有荧光集中斑块。小脑组织切片在颗粒层呈现出丰富的免疫荧光;新生大鼠小脑细胞的解离培养物在颗粒细胞神经元的细胞质和突起中显示出强烈的荧光。数据表明,网格蛋白是一种丰富的蛋白质,明显定位于细胞的细胞质中,并且其分布和排列与肌动蛋白应力纤维明显相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e535/350093/106f9990ed9f/pnas00496-0504-a.jpg

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