Optimization of protein samples for NMR using thermal shift assays

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Kozak, S.; Lercher, L.; Karanth, M.N.; Meijers, R.; Carlomagno, T. et al.: Optimization of protein samples for NMR using thermal shift assays. In: Journal of Biomolecular NMR 64 (2016), Nr. 4, S. 281-289. DOI: http://dx.doi.org/10.1007/s10858-016-0027-z

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Sum total of downloads: 136




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Abstract: 
Maintaining 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. © 2016, The Author(s).
License of this version: CC BY 4.0 Unported
Document Type: Text
Publishing status: publishedVersion
Issue Date: 2016
Appears in Collections:Naturwissenschaftliche Fakultät

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pos. country downloads
total perc.
1 image of flag of Germany Germany 104 76.47%
2 image of flag of United States United States 8 5.88%
3 image of flag of China China 6 4.41%
4 image of flag of Estonia Estonia 1 0.74%
5 image of flag of Denmark Denmark 1 0.74%
6 image of flag of Czech Republic Czech Republic 1 0.74%
7 image of flag of Switzerland Switzerland 1 0.74%
8 image of flag of Canada Canada 1 0.74%
9 image of flag of Australia Australia 1 0.74%
10 image of flag of Argentina Argentina 1 0.74%
    other countries 11 8.09%

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