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中国科学院大学学报 ›› 2013, Vol. 30 ›› Issue (1): 18-23.DOI: 10.7523/j.issn.1002-1175.2013.01.004

• 化学与生命科学 • 上一篇    下一篇

通过对免疫共沉淀技术的优化验证蛋白质弱相互作用

冯晓琴, 徐峰, 王嵬, 刘利新   

  1. 中国科学院研究生院, 北京 100049
  • 收稿日期:2012-02-23 修回日期:2012-03-15 发布日期:2013-01-15
  • 通讯作者: 刘利新
  • 基金资助:

    国家自然科学基金(30670889,30771193)资助

Confirmation of weak protein-protein interactions by optimizing co-immunoprecipitation

FENG Xiao-Qin, XU Feng, WANG Wei, LIU Li-Xin   

  1. Graduate University, Chinese Academy of Sciences, Beijing 100049, China
  • Received:2012-02-23 Revised:2012-03-15 Published:2013-01-15

摘要:

采用单因子法对影响免疫共沉淀结果的各因素进行优化.以hCLP46(human CAP10-like protein46)蛋白和内质网分子伴侣calnexin为例,对相互作用开展研究.通过对细胞裂解液各组分浓度、抗体用量、hCLP46的蛋白量和交联剂DSP因素的优化,验证了hCLP46(human CAP10-like protein46)蛋白和内质网分子伴侣calnexin间的弱相互作用.研究结果为探讨蛋白质之间弱相互作用提供一定的参考价值.

关键词: 免疫共沉淀, hCLP46, calnexin, 影响因素, 蛋白质相互作用

Abstract:

Co-immunoprecipitation is widely used to detect protein-protein interaction in physiological condition. The single-factor method was used to optimize some factors of co-immunoprecipitation, including component concentrations of cell-lysis buffer, antibody dosage, hCLP46 protein quantity, and cross-linking DSP. As a result, the weak interaction between hCLP46 (human CAP10-like protein 46) and ER chaperone calnexin is confirmed. The present work provides an important basis for further study of the protein-protein interaction.

Key words: co-immunoprecipitation(Co-IP), hCLP46, calnexin, optimization factors, protein-protein interaction

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