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兔抗人热激蛋白70样蛋白1多克隆抗体的制备与初步鉴定
引用本文:杨秉芬,李平,李娟,瞿秀华,李晓明,柳晓兰,孙启鸿,曹诚.兔抗人热激蛋白70样蛋白1多克隆抗体的制备与初步鉴定[J].生物技术通讯,2009,20(5):674-676.
作者姓名:杨秉芬  李平  李娟  瞿秀华  李晓明  柳晓兰  孙启鸿  曹诚
作者单位:1. 军事医学科学院,生物工程研究所,北京100850;军事医学科学院,放射与辐射医学研究所,北京100850
2. 军事医学科学院,生物工程研究所,北京100850
3. 军事医学科学院,放射与辐射医学研究所,北京100850
摘    要:目的:制备兔抗人热激蛋白70样蛋白1(HSP70L1)的多克隆抗体并进行初步鉴定。方法:在大肠杆菌中重组表达融合蛋白GST-HSP70L1和His-HSP70L1并纯化;将GST-HSP70L1融合蛋白用于免疫新西兰大耳白兔获得多克隆抗体,用His-HSP70L1对抗血清进行分离纯化,得到抗HSP70L1多克隆抗体,用Western印迹、免疫沉淀对其进行初步鉴定。结果:获得了高表达的GST-HSP70L1和His-HSP70L1重组融合蛋白;纯化获得抗HSP70L1抗体,此抗体可用于Western印迹和免疫沉淀实验。结论:获得了兔抗人HSP70L1的多克隆抗体,为进一步研究HSP70L1的生物功能提供了有用的工具。

关 键 词:热激蛋白70样蛋白1  多克隆抗体  制备  鉴定

Preparation and Preliminary Identification of Rabbit Polyclonal Antibodies Against Human HSP70-Like Protein 1
YANG Bing-Fen,LI Ping,LI Juan,QU Xiu-Hu,LI Xiao-Ming,LIU Xiao-Lan,SUN Qi-Hong,CAO Cheng.Preparation and Preliminary Identification of Rabbit Polyclonal Antibodies Against Human HSP70-Like Protein 1[J].Letters in Biotechnology,2009,20(5):674-676.
Authors:YANG Bing-Fen  LI Ping  LI Juan  QU Xiu-Hu  LI Xiao-Ming  LIU Xiao-Lan  SUN Qi-Hong  CAO Cheng
Institution:1. Beijing Institute of Biotechnology, Beijing 100850; 2. Beijing Institute of Radiation Medicine, Beijing 100850; China)
Abstract:Objective: To preparation and characterize the rabbit polyclonal antibodies against human HSP70-1ike protein 1(HSP70L1). Methods: The fusion recombinant protein GST-HSPTOL1 and His-HSP70L1 were expressed in E.coli. The rabbits were immunized with the purified recombinant protein GST-HSP70L1. The anti-serum was purified with recombinant protein His-HSP70L1. The anti-HSPTOL1 polyclonal antibodies were detected by Western blot and immunoprecipitation. Results: GST-HSP70L1 and His-HSP70L1 were highly expressed in E.coli. Western blot analysis proved the polyclonal antibodies could specifically recognize HSP70L1. Immunoprecipitation confirmed that the polyclonal antibodies could be used to pull down HSP70L1. Conclusion: The rabbit anti-human HSP70L1 polyclonal antibodies were obtained. This study provides useful tools for investigation the biological function of HSP70L1.
Keywords:HSP70-like protein 1  polyclonal antibodies  preparation  identification
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