Optimization of protein samples for NMR using thermal shift assays.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
AbstractMaintaining a stable fold for recombinant proteins is challenging, especially when working with highly purified and concentrated samples at temperatures >20 °C. Therefore, it is worthwhile to screen for different buffer components that can stabilize protein samples. Thermal shift assays or ThermoFluor(®) provide a high-throughput screening method to assess the thermal stability of a sample under several conditions simultaneously. Here, we describe a thermal shift assay that is designed to optimize conditions for nuclear magnetic resonance studies, which typically require stable samples at high concentration and ambient (or higher) temperature. We demonstrate that for two challenging proteins, the multicomponent screen helped to identify ingredients that increased protein stability, leading to clear improvements in the quality of the spectra. Thermal shift assays provide an economic and time-efficient method to find optimal conditions for NMR structural studies.
CitationOptimization of protein samples for NMR using thermal shift assays. 2016, 64 (4):281-9 J. Biomol. NMR
AffiliationHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
JournalJournal of biomolecular NMR
The following license files are associated with this item:
- Creative Commons
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-sa/4.0/
- The combined use of the Thermofluor assay and ThermoQ analytical software for the determination of protein stability and buffer optimization as an aid in protein crystallization.
- Authors: Phillips K, de la Peña AH
- Issue date: 2011 Apr
- Optimization of protein purification and characterization using Thermofluor screens.
- Authors: Boivin S, Kozak S, Meijers R
- Issue date: 2013 Oct
- Analysis of protein stability and ligand interactions by thermal shift assay.
- Authors: Huynh K, Partch CL
- Issue date: 2015 Feb 2
- Thermofluor-based optimization strategy for the stabilization and crystallization of Campylobacter jejuni desulforubrerythrin.
- Authors: Santos SP, Bandeiras TM, Pinto AF, Teixeira M, Carrondo MA, Romão CV
- Issue date: 2012 Feb
- How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project.
- Authors: Bruce D, Cardew E, Freitag-Pohl S, Pohl E
- Issue date: 2019 Feb 11