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利用抗小鼠 ADP 核糖聚合物的多克隆抗体,通过狭缝印迹和 Western 印迹免疫检测法在体内检测和定量小鼠脾脏和肝脏组织中多聚 ADP 核糖基化的细胞蛋白。

Detection and quantification of poly-ADP-ribosylated cellular proteins of spleen and liver tissues of mice in vivo by slot and Western blot immunoprobing using polyclonal antibody against mouse ADP-ribose polymer.

作者信息

Sharan R N, Devi B Jaylata, Humtsoe J O, Saikia Jyoti R, Kma L

机构信息

Radiation and Molecular Biology Unit, Department of Biochemistry, North-Eastern Hill University, Umshing, Shillong, India.

出版信息

Mol Cell Biochem. 2005 Oct;278(1-2):213-21. doi: 10.1007/s11010-005-7588-6.

Abstract

Poly-ADP-ribosylation (PAR) of cellular proteins has been shown to have decisive roles in diverse cellular functions including carcinogenesis. There are indications that metabolic level of poly-ADP-ribosylated cellular proteins might indicate carcinogenesis and, therefore, could be potentially used in cancer screening program. Keeping in mind the limitations of currently available assays of cellular PAR, a new assay is being reported that measures the metabolic level of poly-ADP-ribosylated cellular proteins. The ELISA based slot and Western blot immunoassay used polyclonal antibody against natural, heterogeneous ADP-ribose polymers. It could be successfully employed to qualitatively and quantitatively assay metabolic levels of poly-ADP-ribosylated proteins of spleen and liver tissues of normal mice or mice exposed to dimethylnitrosamine for up to 8 weeks; potentially PAR of cellular proteins could be assayed in any tissue or biopsy. Implications of the results in cancer screening program have been discussed.

摘要

细胞蛋白的多聚ADP核糖基化(PAR)已被证明在包括致癌作用在内的多种细胞功能中起决定性作用。有迹象表明,多聚ADP核糖基化细胞蛋白的代谢水平可能表明致癌作用,因此有可能用于癌症筛查项目。考虑到目前可用的细胞PAR检测方法的局限性,本文报道了一种新的检测方法,该方法可测量多聚ADP核糖基化细胞蛋白的代谢水平。基于酶联免疫吸附测定(ELISA)的狭缝印迹和蛋白质免疫印迹法使用了针对天然、异质性ADP核糖聚合物的多克隆抗体。该方法可成功用于定性和定量检测正常小鼠或暴露于二甲基亚硝胺长达8周的小鼠脾脏和肝脏组织中多聚ADP核糖基化蛋白的代谢水平;潜在地,细胞蛋白的PAR可在任何组织或活检样本中进行检测。本文还讨论了这些结果在癌症筛查项目中的意义。

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