Preparation of polyclonal antibodies against the Drosophila deacetylases SIRT 6 and SIRT 7

多克隆抗体 重组DNA 生物 抗血清 抗体 免疫印迹 癌变 乙酰化 果蝇属(亚属) SIRT6型 锡尔图因 分子生物学 细胞生物学 遗传学 基因
作者
Zhenhao Wei,Jiejie Zhu,Yafei Cai,Ta Liu,MA Xiang-hua,Xiaodie Feng,Yaoyao Wang,Yushan Li,Wei Zhang
出处
期刊:Protein Expression and Purification [Elsevier BV]
卷期号:211: 106338-106338
标识
DOI:10.1016/j.pep.2023.106338
摘要

SIRT6 and SIRT7, as members of the sirtuins family, are indispensable for the growth and development of Drosophila. They play crucial roles in maintaining genome stability, regulating metabolic senescence, and controlling tumorigenesis. To investigate their involvement in the Drosophila life cycle, we focused on describing the expression and purification of recombinant Drosophila SIRT6 and SIRT7 proteins. Subsequently, these proteins were utilized for generating polyclonal antibodies against Drosophila SIRT6 and SIRT7. The recombinant expression plasmid was introduced into E. coli cells to enable the production of SIRT6 and SIRT7 proteins. Following immunizations of New Zealand white rabbits and guinea pigs with the recombinant proteins as antigens, specific polyclonal antisera against both proteins were obtained. After purification, the specificity of SIRT6 and SIRT7 was confirmed using ELISA and western blot analyses, demonstrating strong specificity. These antibodies hold promise for the development of detection assays required for further research.

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