基于串联杂合泛素结合结构域探针的多聚泛素化信号免疫组织化学检测技术研究  

Immunohistochemical detection technology for polyubiquitination signals based on the tandem hybrid ubiquitin-binding domain probe

作　　者：黄帅 肖伟弟 李衍常 徐平 HUANG Shuai;XIAO Wei-di;LI Yan-chang;XU Ping(State Key Laboratory of Proteomics,National Center for Protein Sciences,Institute of Lifeomics,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 102206,China)

机构地区：[1]军事科学院军事医学研究院生命组学研究所,国家蛋白质科学中心,蛋白质组学国家重点实验室,北京102206

出　　处：《军事医学》2023年第3期205-209,共5页Military Medical Sciences

基　　金：科技部重点研发计划(2020YFE0202200,2017YFA0505000);国家自然科学基金项目(32071431,31901037)。

摘　　要：目的提高串联杂合泛素结合结构域(ThUBD)探针检测甲醛固定石蜡包埋(FFPE)切片中多聚泛素化信号的灵敏度。方法利用微波炉加热方法,采用不同盐离子种类和pH缓冲液进行抗原修复,在有无高浓度还原剂条件下,用ThUBD探针检测FFPE切片中的多聚泛素化信号。结果在pH 9.5 EDTA缓冲液条件下,加入50 mmol/L DTT可检测到FFPE切片中明显的多聚泛素化信号。结论pH 9.5 EDTA缓冲液中加50 mmol/L DTT有利于多聚泛素链的暴露与复原,显著提升ThUBD探针用于免疫组织化学检测的灵敏度。Objective To improve the sensitivity of the tandem hybrid ubiquitin-binding domain(ThUBD)probe for detecting polyubiquitination signals in formaldehyde-fixed paraffin-embedded(FFPE)sections.Methods Buffer solutions with different salt ions and pH values were used for antigen retrieval via microwave.The polyubiquitination signals in FFPE sections were detected by the ThUBD probe with or without any reducing agent of a high concentration.Results Addition of 50 mmol/L DTT in pH 9.5 EDTA buffer solutions enhanced polyubiquitination signals in FFPE sections.Conclusion pH 9.5 EDTA buffer solutions plus 50 mmol/L DTT can facilitate the exposure and structure recovery of polyubiquitin chains,and significantly improve the sensitivity of the ThUBD probe for immunohistochemical detection.

关 键 词：甲醛固定石蜡包埋切片 抗原修复 多聚泛素化信号检测 免疫组织化学 串联杂合泛素结合结构域探针 

分 类 号：R392.11[医药卫生—免疫学]