Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: Jul 20, 2026

How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project
07:22

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

Published on: February 11, 2019

Thermofluor-based high-throughput stability optimization of proteins for structural studies.

Ulrika B Ericsson1, B Martin Hallberg, George T Detitta

  • 1Department of Biochemistry and Biophysics, Stockholm University, Stockholm SE-109 51, Sweden.

Analytical Biochemistry
|September 12, 2006
PubMed
Summary

Thermofluor screening rapidly identifies optimal protein formulations and buffers for crystallization. This high-throughput method improves protein crystallization success rates by identifying stabilizing additives and ligands.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Pleiotropic effects of BET inhibition broadly boost tumor immunogenicity to CD8<sup><b>+</b></sup> T cells.

Oncoimmunology·2026
Same author

Hybrid Extracellular Vesicles for Efficient Loading and Functional Delivery of mRNA.

Journal of extracellular vesicles·2025
Same author

Correction to "Intracellular Binding of Novel Fluorinated Compounds to Carbonic Anhydrase Isoforms Explored by In-Cell <sup>19</sup>F NMR".

Journal of medicinal chemistry·2025
Same author

Loop-mediated regulation and base flipping drive RNA cleavage by human mitochondrial PNPase.

Nucleic acids research·2025
Same author

Intracellular Binding of Novel Fluorinated Compounds to Carbonic Anhydrase Isoforms Explored by In-Cell <sup>19</sup>F NMR.

Journal of medicinal chemistry·2025
Same author

The herpes simplex origin-binding protein: mechanisms for sequence-specific DNA binding and dimerization revealed by Cryo-EM.

Nucleic acids research·2025

Area of Science:

  • Structural Biology
  • Biochemistry
  • Protein Crystallization

Background:

  • Protein structure determination is crucial for understanding biological function.
  • Protein sample properties like homogeneity, stability, and solubility significantly impact crystallization success.
  • Optimizing these properties is essential for improving crystallization yields.

Purpose of the Study:

  • To apply the thermofluor method as a high-throughput approach for identifying optimal protein formulations for crystallization.
  • To rapidly identify optimal stabilizing buffer compositions for individual proteins.
  • To identify potential ligands that can enhance subsequent crystallization trials.

Main Methods:

  • Thermofluor screening was employed to determine thermally induced melting points.

More Related Videos

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O
03:09

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O

Published on: August 9, 2024

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
13:26

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry

Published on: September 13, 2014

Related Experiment Videos

Last Updated: Jul 20, 2026

How to Stabilize Protein: Stability Screens for Thermal Shift Assays and Nano Differential Scanning Fluorimetry in the Virus-X Project
07:22

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

Published on: February 11, 2019

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O
03:09

Utilizing Thermal Shift Assay to Probe Substrate Binding to Selenoprotein O

Published on: August 9, 2024

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
13:26

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry

Published on: September 13, 2014

  • Melting points were measured for 25 Escherichia coli proteins in various buffers and with additives.
  • Crystallization trials were conducted using stabilizing and destabilizing additives identified via thermofluor screening.

    • Main Results:

    • The thermofluor method efficiently identified optimal buffer compositions and stabilizing additives.
    • A twofold increase in crystallization leads was observed when using stabilizing additives identified by thermofluor.
    • The method facilitated the identification of ligands that can be used in crystallization.

    Conclusions:

    • Thermofluor is an efficient, high-throughput method for optimizing protein formulations for crystallization.
    • Identifying protein properties predictive of crystallizability using thermofluor enhances success rates.
    • This approach accelerates the process of obtaining high-quality protein crystals for structural studies.