基于全内反射荧光的单分子检测对组蛋白修饰酶进行多重检测。

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

作者信息

Ma Fei, Liu Meng, Wang Zi-yue, Zhang Chun-yang

机构信息

College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals, Shandong Normal University, Jinan 250014, China.

出版信息

Chem Commun (Camb). 2016 Jan 21;52(6):1218-21. doi: 10.1039/c5cc08797j.

DOI:10.1039/c5cc08797j

PMID:26608941

Abstract

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.

摘要

我们通过将基于抗体的荧光标记与基于全内反射荧光（TIRF）的单分子检测相结合，开发了一种用于多重检测组蛋白修饰酶（HMEs）的灵敏且具选择性的方法。该方法具有出色的特异性和高灵敏度，对组蛋白乙酰转移酶GcN5的检测限为21 pM，对组蛋白甲基转移酶G9a的检测限为12 pM，并且它也可用于筛选HME抑制剂。