期刊
NATURE PROTOCOLS
卷 11, 期 5, 页码 882-894出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2016.044
关键词
-
资金
- Deutsche Forschungsgemeinschaft (DFG) through International Research Training Group [1830]
- Stiftung Rheinland-Pfalz fur Innovation
- Intramural Research Program of the National Institute of Biomedical Imaging and Bioengineering, US National Institutes of Health
Isothermal titration calorimetry (ITCTC) is a powerful and widely used method to measure the energetics of macromolecular interactions by recording a thermogram of differential heating power during a titration. However, traditional ITCTC analysis is limited by stochastic thermogram noise and by the limited information content of a single titration experiment. Here we present a protocol for bias-free thermogram integration based on automated shape analysis of the injection peaks, followed by combination of isotherms from different calorimetric titration experiments into a global analysis, statistical analysis of binding parameters and graphical presentation of the results. This is performed using the integrated public-domain software packages NITPTPIC, SESEDPHATAT and GUSSUSSUSSI. The recently developed low-noise thermogram integration approach and global analysis allow for more precise parameter estimates and more reliable quantification of multisite and multicomponent cooperative and competitive interactions. Titration experiments typically take 1-2.5 h each, and global analysis usually takes 10-20 min.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据