Using Differential Scanning Fluorimetry (DSF) to Detect Ligand Binding with Purified Protein

Xiaohui Li; Chunhua Zhang

doi:10.1007/978-1-0716-0954-5_16

Using Differential Scanning Fluorimetry (DSF) to Detect Ligand Binding with Purified Protein

Methods Mol Biol. 2021:2213:183-186. doi: 10.1007/978-1-0716-0954-5_16.

Authors

Xiaohui Li^{1

2}, Chunhua Zhang³

Affiliations

¹ Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN, USA.
² Purdue Center for Plant Biology, Purdue University, West Lafayette, IN, USA.
³ Department of Botany and Plant Pathology, Center for Plant Biology, Purdue University, West Lafayette, IN, USA. zhang150@purdue.edu.

PMID: 33270203
DOI: 10.1007/978-1-0716-0954-5_16

Abstract

Differential scanning fluorimetry (DSF) can be used to detect the binding of a small molecule ligand to a purified target protein. Upon binding with certain ligands, the protein can be stabilized from thermal denaturation. DSF uses a fluorescent dye and Real-Time PCR instrument to detect the unfolding process of proteins during thermal denaturation. The experiment can be set up and finished in 1 day once the purified protein is available.

Keywords: Differential scanning fluorimetry; Ligand binding; Purified protein; Real-Time PCR instrument.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Data Analysis
Fluorometry / methods*
Ligands
Proteins / isolation & purification*
Temperature

Substances

Ligands
Proteins