[Preparation and identification of rabbit anti-cyclin dependent kinase 6 (CDK6) antibodies]

科研成果详情

题名	[Preparation and identification of rabbit anti-cyclin dependent kinase 6 (CDK6) antibodies]
作者	Xiaoxian Ye1; Haiyan Dong2; Yu Wang 1; Zhengzhen Chen1; Junwei Li 1; Yubing Wei1; Lifang Zhang 3
发表日期	2023-08
发表期刊	Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 影响因子和分区
语种	中文
原始文献类型	English Abstract ; Journal Article
摘要	Objective To prepare and identify rabbit anti-cyclin dependent kinase 6 (CDK6) antibody. Methods The recombinant pET21a (+)/CDK6 was successfully constructed, then the recombinant plasmid was transformed into E.coli BL21 (DE3) competent cells and was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) for protein expression, which was detected by SDS-PAGE and Western blot analysis. The expressed protein was purified by nickel-chelating nitrilotriacetic acid (Ni-NTA) agarose and then analyzed by SDS-PAGE. Japanese white rabbits were immunized with purified CDK6 protein for many times every two weeks. The blood was collected at 0, 2, 4 and 6 weeks after immunization, and serum was separated from blood. The titer was detected by indirect ELISA. Western blot analysis, immunofluorescence assay and immunohistochemistry were employed to determine the specificity. Results High purity CDK6 protein and high specificity of rabbit anti-CDK6 antibody were successfully prepared. The titer of CDK6 rabbit serum antibody reached 1:30 000 after immunization, which could specifically recognize the CDK6 protein expressed in cervical cancer cell line and cervical cancer tissues. Conclusion The high titer and specificity of rabbit anti-CDK6 antibody is successfully prepared.
ISSN	1007-8738
卷号	39 期号:8
收录类别	PUBMED
URL	查看原文
PubMed ID	37515341
文献类型	期刊论文
条目标识符	https://kms.wmu.edu.cn/handle/3ETUA0LF/181758
专题	基础医学院（机能实验教学中心）仁济学院
作者单位	1.Department of Anatomy, School of Basic Medicine, Wenzhou Medical University Renji College, Wenzhou 325035, China.; 2.Institute of Molecular Viruses and Immunology, Wenzhou Medical University Renji College, Wenzhou 325035, China.; 3.Institute of Molecular Viruses and Immunology, Wenzhou Medical University Renji College, Wenzhou 325035, China. *Corresponding author, E-mail: zhanglifang08@126.com.
第一作者单位	基础医学院（机能实验教学中心）
第一作者的第一单位	基础医学院（机能实验教学中心）
推荐引用方式 GB/T 7714	Xiaoxian Ye,Haiyan Dong,Yu Wang,et al. [Preparation and identification of rabbit anti-cyclin dependent kinase 6 (CDK6) antibodies][J]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology,2023,39(8).
APA	Xiaoxian Ye., Haiyan Dong., Yu Wang., Zhengzhen Chen., Junwei Li., ... & Lifang Zhang. (2023). [Preparation and identification of rabbit anti-cyclin dependent kinase 6 (CDK6) antibodies]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, 39(8).
MLA	Xiaoxian Ye,et al."[Preparation and identification of rabbit anti-cyclin dependent kinase 6 (CDK6) antibodies]".Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 39.8(2023).