Rivista
/
Biochimica
/
How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project
JoVE Journal
Biochimica
This content is Free Access.
JoVE Journal Biochimica
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

•

07:22 min

•

February 11, 2019

•

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

Capitoli

  • 00:04Titolo
  • 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

Traduzione automatica

Traduzione automatica

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

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

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

check_url/it/58666?article_type=v

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro
check_url/it/58666?article_type=v

Differential Scanning Calorimetry — A Method for Assessing the Thermal Stability and Conformation of Protein Antigen
check_url/it/58666?article_type=v

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
check_url/it/58666?article_type=v

Protein-tRNA Agarose Gel Retardation Assays for the Analysis of the N6-threonylcarbamoyladenosine TcdA Function
check_url/it/58666?article_type=v

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
check_url/it/58666?article_type=v

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator
check_url/it/58666?article_type=v

How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
check_url/it/58666?article_type=v

Single Liposome Measurements for the Study of Proton-Pumping Membrane Enzymes Using Electrochemistry and Fluorescent Microscopy
check_url/it/58666?article_type=v

Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase
check_url/it/58666?article_type=v

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

Read Article