Multiplex detection of histone-modifying enzymes by total internal reflection fluorescence-based single-molecule detection

被引:50
作者
Ma, Fei [1 ,2 ]
Liu, Meng [1 ]
Wang, Zi-yue [1 ]
Zhang, Chun-yang [1 ,2 ]
机构
[1] Shandong Normal Univ, Coll Chem Chem Engn & Mat Sci, Collaborat Innovat Ctr Functionalized Probes Chem, Key Lab Mol & Nano Probes,Minist Educ,Shandong Pr, Jinan 250014, Peoples R China
[2] Chinese Acad Sci, Shenzhen Inst Adv Technol, Single Mol Detect & Imaging Lab, Shenzhen 518055, Peoples R China
基金
中国国家自然科学基金;
关键词
PROTEIN-PHOSPHORYLATION; DNA-REPLICATION; ASSAY; ACETYLATION; METHYLTRANSFERASES; GCN5; H3; ACETYLTRANSFERASES; SPECIFICITY; COMPLEXES;
D O I
10.1039/c5cc08797j
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We develop a sensitive and selective method for the multiplex detection of histone-modifying enzymes (HMEs) through the integration of antibody-based fluorescence labeling with total internal reflection fluorescence (TIRF)-based single-molecule detection. This method exhibits excellent specificity and high sensitivity with a detection limit of 21 pM for histone acetyltransferase GcN5 and 12 pM for histone methyltransferase G9a, and it can be applied for the screening of HME inhibitors as well.
引用
收藏
页码:1218 / 1221
页数:4
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