Label-Free Determination of the Dissociation Constant of Small Molecule-Aptamer Interaction by Isothermal Titration Calorimetry.

• 发表时间: 2016
• 来源期刊: Methods Mol Biol
• DOI: 10.1007/978-1-4939-3197-2_9
• PMID: 26552820
• 文章类型: Journal Article
• 作者列表: Vogel M,Suess B

关键词: Aptamer; ITC; In vitro transcription; K d; RNA; Small molecule

Mesh: Aptamers, Nucleotide / chemistry genetics metabolism; Calorimetry / methods; In Vitro Techniques; Mutation; RNA / chemistry genetics metabolism; Thermodynamics; Titrimetry / methods; Transcription, Genetic

Abstract

Isothermal titration calorimetry (ITC) is a powerful label-free technique to determine the binding constant as well as thermodynamic parameters of a binding reaction and is therefore well suited for the analysis of small molecule-RNA aptamer interaction. We will introduce you to the method and present a protocol for sample preparation and the calorimetric measurement. A detailed note section will point out useful tips and pitfalls.