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How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project
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Biochemistry
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JoVE Journal Biochemistry
How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project

How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project

DOI:
10.3791/58666-v

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07:22 min

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February 11, 2019

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, , ,
1Department of Biosciences,Durham University, 2Department of Chemistry,Durham University

Chapters

  • 00:04Title
  • 00:27Preparation of Protein Sample
  • 01:31Performing a Thermal Shift Assay (TSA) Experiment
  • 03:22Preparing and Programing a Nano Differential Scanning Fluorimetry (nanoDSF) Experiment
  • 05:35Results: Analysis of the Temperature at the Inflection Point of the Unfolding Transition (Tm) in Lysozyme
  • 06:55Conclusion

Summary

Automatic Translation

Automatic Translation

A protocol is presented to rapidly test the thermal stability of proteins in a variety of conditions through thermal shift assays and nano differential scanning fluorimetry. Buffer systems, salts and additives, together comprising three unique stability screens, are assayed with proteins to identify suitable buffers for functional and structural studies.

Tags

Keywords: Protein StabilizationThermal Shift AssayNano Differential Scanning FluorimetryProtein ChemistryDrug DiscoveryStability ScreenProtein ThermostabilityProtein Ligand BindingHigh-throughputSYPRO Orange DyeRT-PCR SystemMelt Curve Analysis

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