激光生物学报2012，Vol.21Issue(1)：56-59,4.DOI:10.3969/j.issn.1007-7146.2012.01.012
斑马鱼基因ATF4多克隆抗体的制备
Preparation of Polyclonal Antibody of ATF4 in Zebrafish
摘要
Abstract
ATF4 , a member of the ATF/CREB transcription factor family which contains a bZIP domain, plays a important role in the embryonic development, cell proliferation and differentiation. Therefore, it is advisable to prepare an antibody for the heart development research in Zerbrafish. In this research, a ATF4-specific, hydrophilic nucleotide sequence (about 1 017 bp) was selected by using bioinformatics methods, and the sequence was amplified by PCR, the PCR product was inserted into the prokaryotic expression vector pET-28a and then transformed into Rosetta cells. After the sequence result of the recombinant plasmid pET-28a-ATF4 is verified to be right, the fusion protein was induced to express in Rosetta cells by IPTG. The mouse was immunized by injecting appropriate amount of fusion protein, and the ATF4 polyclonal antibody was obtained. The antibody' s titer and specification was identified. Our results showed that the ATF4 polyclonal antibody was practical and available for western-blotting analysis and immunohisto-chemistry.关键词
ATF4/融合蛋白/多克隆抗体Key words
ATF4/ fusion protein/ polyclonal antibody分类
生物科学引用本文复制引用
李发祥,杨荣,陈婷芳,唐雄卓,赵碧峰,江志钢,张义,李永青,吴秀山..斑马鱼基因ATF4多克隆抗体的制备[J].激光生物学报,2012,21(1):56-59,4.基金项目
国家自然科学基金资助项目(30930054) （30930054）
