articleCurrent Protocols in Protein ScienceFeb 1, 2015GREEN OA

Analysis of Protein Stability and Ligand Interactions by Thermal Shift Assay

KHKathy HuynhCLCarrie L. Partch

University of California, Santa Cruz

PubMed
Indexed incrossrefpubmed

Abstract

Purification of recombinant proteins for biochemical assays and structural studies is time-consuming and presents inherent difficulties that depend on the optimization of protein stability. The use of dyes to monitor thermal denaturation of proteins with sensitive fluorescence detection enables rapid and inexpensive determination of protein stability using real-time PCR instruments. By screening a wide range of solution conditions and additives in a 96-well format, the thermal shift assay easily identifies conditions that significantly enhance the stability of recombinant proteins. The same approach can be used as an initial low-cost screen to discover new protein-ligand interactions by capitalizing on…

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Topics & keywords

Topics
Keywords
  • Recombinant DNA
  • Thermal stability
  • Chemistry
  • Ligand (biochemistry)
  • Denaturation (fissile materials)
  • Protein stability
  • Fluorescence
  • Target protein
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