{"title":"Generation of Rat Monoclonal Antibody for Mouse Nucleolin by Immunization of Ferroptosis-Induced Hepa 1-6 Cells.","authors":"Chikako Yokoyama, Sho Kobayashi, Yumi Harada, Yuki Nishino, Junichi Fujii, Taro Tachibana","doi":"10.1089/mab.2022.0005","DOIUrl":null,"url":null,"abstract":"<p><p>Nucleolin is a multifunctional phosphoprotein that is ubiquitously distributed in the nucleus, nucleoplasm, cytoplasm, and cell membrane. The principal functions of nucleolin involve DNA and RNA metabolism, gene transcription and translation, ribosome biogenesis, and mRNA stability. Ferroptosis is a type of regulated cell death that is characterized by the iron-dependent accumulation of lipid peroxidation products. In a previous study, we produced monoclonal antibodies (mAbs) against lysates prepared from ferroptosis-induced Hepa 1-6 cells. In this study, we describe one of those rat mAbs, 4B5, which was generated against mouse nucleolin. This mAb was useful in immunofluorescence staining, immunoblotting, and immunoprecipitation experiments, and was confirmed to recognize endogenous nucleolin in mouse cell lines and tissues. We anticipate that mAb 4B5 will be useful for functional analyses of nucleolin.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"255-259"},"PeriodicalIF":0.0000,"publicationDate":"2022-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/mab.2022.0005","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/10/20 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0
Abstract
Nucleolin is a multifunctional phosphoprotein that is ubiquitously distributed in the nucleus, nucleoplasm, cytoplasm, and cell membrane. The principal functions of nucleolin involve DNA and RNA metabolism, gene transcription and translation, ribosome biogenesis, and mRNA stability. Ferroptosis is a type of regulated cell death that is characterized by the iron-dependent accumulation of lipid peroxidation products. In a previous study, we produced monoclonal antibodies (mAbs) against lysates prepared from ferroptosis-induced Hepa 1-6 cells. In this study, we describe one of those rat mAbs, 4B5, which was generated against mouse nucleolin. This mAb was useful in immunofluorescence staining, immunoblotting, and immunoprecipitation experiments, and was confirmed to recognize endogenous nucleolin in mouse cell lines and tissues. We anticipate that mAb 4B5 will be useful for functional analyses of nucleolin.
核蛋白是一种多功能磷蛋白，广泛分布于细胞核、核质、细胞质和细胞膜中。核蛋白的主要功能涉及 DNA 和 RNA 代谢、基因转录和翻译、核糖体生物发生和 mRNA 稳定。铁突变是一种调节性细胞死亡，其特点是脂质过氧化产物的铁依赖性积累。在之前的一项研究中，我们制备了针对铁凋亡诱导的 Hepa 1-6 细胞裂解物的单克隆抗体（mAbs）。在本研究中，我们介绍了其中一种针对小鼠核苷酸的大鼠 mAb 4B5。这种 mAb 可用于免疫荧光染色、免疫印迹和免疫沉淀实验，并被证实能识别小鼠细胞系和组织中的内源性核素。我们预计 mAb 4B5 将有助于对 nucleolin 进行功能分析。
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