Chen Miao, Yang Jin-Ju, Li Shu-Zhen, Liu Xiao-Lan, Liu Ying, Zhang Lin-Jie, Gao Jian-En, Sun Qi-Hong
College of Preclinical Medicine, Anhui Medical University, Hefei 230032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jul;24(7):699-702.
To preparation and characterize the rabbit polyclonal antibodies against human PON2 (paraoxonase-2).
A fragment of human PON2 gene which was of low homology with rabbits but of higher hydrophilicity and immunogenicity was selected for recombinant expression in prokaryotic expression system. The rabbits were immunized with the purified GST fusion protein 3 times. The specificity and sensitivity of the anti-human PON2 polyclonal antibodies were detected by Western blot and indirect immunofluorescence.
The GST-PON2 fusion protein was highly expressed in Ecoli with a molecular weight of 46 kDa. Western blot analysis proved the rabbit polyclonal antibodies could specifically recognize 39 kDa native PON2 protein expressed in several cells and tissues, such as HeLa cells, U937 cells, and human liver tissue. Indirect immunofluorescence analysis confirmed that PON2 protein was located in the cytoplasm of SY5Y cells.
The rabbit polyclonal antibodies against human PON2 can specifically recognize natural protein expressed in human cells and tissues, Which can be used for further study and clinical detection of human PON2.
制备并鉴定抗人对氧磷酶2（PON2）的兔多克隆抗体。
选择一段与人PON2基因同源性低但亲水性和免疫原性较高的片段，在原核表达系统中进行重组表达。用纯化的GST融合蛋白对兔进行3次免疫。通过蛋白质免疫印迹法（Western blot）和间接免疫荧光法检测抗人PON2多克隆抗体的特异性和敏感性。
GST-PON2融合蛋白在大肠杆菌中高表达，分子量为46 kDa。蛋白质免疫印迹分析证明，兔多克隆抗体能够特异性识别在多种细胞和组织（如HeLa细胞、U937细胞和人肝组织）中表达的39 kDa天然PON2蛋白。间接免疫荧光分析证实，PON2蛋白位于SY5Y细胞的细胞质中。
抗人PON2兔多克隆抗体能够特异性识别在人细胞和组织中表达的天然蛋白，可用于人PON2的进一步研究和临床检测。
