An effective thiol-reactive probe for differential scanning fluorimetry with a standard real-time polymerase chain reaction device

摘要

Differential scanning fluorimetry (DSF) is used to assess protein stability, transition states, or the K-d values of various ligands, drug molecules, and antibodies. All fluorescent probes published to date either are incompatible with hydrophobic proteins/ligands, precluding analyses of transmembrane or membrane-associated proteins, or have excitation and detection wavelengths outside the range of real-time polymerase chain reaction (RT-PCR) machines, necessitating the use of dedicated devices. Here, we describe a thiol-reactive probe, BODIPY FL L-cystine (BFC), to overcome both of these shortcomings. The probe supports an inexpensive application of DSF measurements suitable for detection with standard RT-PCR machines in a hydrophilic or hydrophobic environment. (C) 2016 Elsevier Inc. All rights reserved.