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通过单克隆抗体分析S100相关钙结合蛋白pEL98在成纤维细胞中的细胞定位及其组织分布。

Cellular localization of pEL98 protein, an S100-related calcium binding protein, in fibroblasts and its tissue distribution analyzed by monoclonal antibodies.

作者信息

Takenaga K, Nakamura Y, Sakiyama S

机构信息

Division of Chemotherapy, Chiba Cancer Center Research Institute, Japan.

出版信息

Cell Struct Funct. 1994 Jun;19(3):133-41. doi: 10.1247/csf.19.133.

Abstract

Two monoclonal antibodies (named mAb3G4 and mAb4A2) against recombinant pEL98 protein have been produced. The specificity of the mAbs was verified by immunoblot and immunoprecipitation analyses. To determine cellular localization of the pEL98 protein in NIH3T3 and 3Y1 cells, indirect immunofluorescent staining was performed using mAb4A2. The results showed that most of the pEL98 protein existed diffusely in the cytoplasm, while a part of the protein existed along actin stress fibers in both cells. The expression of pEL98 protein in various murine and human tissues and cells was also examined by immunoblot analysis using mAb3G4. The results demonstrated that the pEL98 protein was expressed in bone marrow, spleen, peritoneal macrophages and human lymphocytes, with the highest amount found in peritoneal macrophages.

摘要

已制备出两种针对重组pEL98蛋白的单克隆抗体(分别命名为mAb3G4和mAb4A2)。通过免疫印迹和免疫沉淀分析验证了单克隆抗体的特异性。为了确定pEL98蛋白在NIH3T3和3Y1细胞中的细胞定位,使用mAb4A2进行了间接免疫荧光染色。结果表明,大部分pEL98蛋白弥散存在于细胞质中,而在这两种细胞中,一部分蛋白沿着肌动蛋白应力纤维存在。还使用mAb3G4通过免疫印迹分析检测了pEL98蛋白在各种小鼠和人类组织及细胞中的表达。结果表明,pEL98蛋白在骨髓、脾脏、腹腔巨噬细胞和人类淋巴细胞中表达,在腹腔巨噬细胞中含量最高。

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