Nakamura Hideaki, Asai Azusa, Maruyama Mitsuo, Sugimoto Masataka
Department of Mechanism of Aging, National Institute for Longevity Sciences, NCGG, Aichi, Japan.
Hybridoma (Larchmt). 2010 Feb;29(1):7-11. doi: 10.1089/hyb.2009.0057.
The hematopoietic zinc finger protein, Hzf, is induced in response to DNA damage or by Arf tumor suppressor in a p53-dependent manner. Recent studies have revealed that Hzf is an RNA-binding protein that regulates localization and translation of specific mRNA. The RNA-binding activity of Hzf is required for the functions of cerebellar purkinje cells and adipocytes, although their molecular mechanisms underlying the mRNA regulation largely remain unknown. To further investigate the molecular function of Hzf, we raised two rat monoclonal antibodies (MAb) against a peptide corresponding to the C-terminal region of the mouse/human Hzf protein. Both MAbs reacted with the native protein expressed in mammalian cells, and were highly efficient in detecting endogenous Hzf by immunoblotting, immunoprecipitation, and immunofluorescence. These MAbs should therefore be useful for further analysis of molecular functions of the Hzf protein and for identification of Hzf-binding proteins.
造血锌指蛋白Hzf在DNA损伤应答中或由Arf肿瘤抑制因子以p53依赖的方式诱导产生。最近的研究表明，Hzf是一种RNA结合蛋白，可调节特定mRNA的定位和翻译。尽管Hzf在mRNA调节方面的分子机制很大程度上仍不清楚，但其RNA结合活性是小脑浦肯野细胞和脂肪细胞功能所必需的。为了进一步研究Hzf的分子功能，我们制备了两种大鼠单克隆抗体（MAb），它们针对与小鼠/人类Hzf蛋白C末端区域对应的肽段。这两种单克隆抗体均与哺乳动物细胞中表达的天然蛋白发生反应，并且在通过免疫印迹、免疫沉淀和免疫荧光检测内源性Hzf方面效率很高。因此，这些单克隆抗体对于进一步分析Hzf蛋白的分子功能以及鉴定Hzf结合蛋白应该是有用的。
