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使用COOH末端肽制备抗大鼠锰超氧化物歧化酶单克隆抗体。

Preparation of a monoclonal antibody against rat MnSOD, using a COOH-terminal peptide.

作者信息

Ria F, Landriscina M, Galeotti T

机构信息

Institute of General Pathology, Catholic University, Rome, Italy.

出版信息

Biochem Biophys Res Commun. 1993 Sep 15;195(2):697-703. doi: 10.1006/bbrc.1993.2101.

Abstract

In the present paper we report the production of a monoclonal antibody against rat MnSOD, a supposed tumor-suppressor protein, using a purified synthetic peptide encompassing amino acids 184-198 to immunize mice, without conjugation to a carrier. The resulting antibody is able to recognize the native form of the protein, since it can immunoprecipitate the MnSOD activity in rat liver homogenate. In Western blot studies, the antibody recognizes a protein of 24 KD M(r), whose concentration varies according to the MnSOD activity and it apparently recognizes also human and mouse MnSODs. The protocol of immunization gives high yield of secreting lines. This monoclonal antibody will allow the detection of structural and functional alterations of MnSOD.

摘要

在本论文中,我们报道了一种针对大鼠锰超氧化物歧化酶(一种假定的肿瘤抑制蛋白)的单克隆抗体的制备。我们使用包含184 - 198位氨基酸的纯化合成肽免疫小鼠,且未与载体偶联。所得抗体能够识别该蛋白的天然形式,因为它可以免疫沉淀大鼠肝脏匀浆中的锰超氧化物歧化酶活性。在蛋白质印迹研究中,该抗体识别一种分子量为24 KD的蛋白质,其浓度随锰超氧化物歧化酶活性而变化,并且它显然也能识别人类和小鼠的锰超氧化物歧化酶。免疫方案产生了高产率的分泌细胞系。这种单克隆抗体将有助于检测锰超氧化物歧化酶的结构和功能改变。

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