title: Time-Dependent Protein Thermostability Assay.
abstract: Membrane protein purification often yields rather unstable proteins impeding functional and structural protein characterization. Low protein stability also leads to low purification yields as a result of protein degradation, aggregation, precipitation, and folding instability. It is often required to optimize buffer conditions through numerous iterations of trial and error to improve the homogeneity, stability, and solubility of the protein sample demanding high amounts of purified protein. Therefore we have set up a fast, simple, and high-throughput time-dependent thermostability-based assay at low protein cost to identify protein stabilizing factors to facilitate the handling and characterization of membrane proteins by subsequent structural and functional studies.
authors.affiliation: Department of Cellular and Molecular Medicine, Laboratory of Cellular Transport Systems, KU Leuven, Herestraat 49, 802, B3000, Leuven, Belgium.
authors.forename: Ilse
authors.initials: I
authors.lastname: Vandecaetsbeek
authors.affiliation: Department of Cellular and Molecular Medicine, Laboratory of Cellular Transport Systems, KU Leuven, Herestraat 49, 802, B3000, Leuven, Belgium. Peter.Vangheluwe@med.kuleuven.be.
authors.forename: Peter
authors.initials: P
authors.lastname: Vangheluwe
mesh_terms: D001681:Biological Assay / Q000379:methods*; D002021:Buffers; D002846:Chromatography, Affinity / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D055550:Protein Stability; D059748:Proteolysis; D012995:Solubility; D013696:Temperature
chemical_list: D002021:Buffers; D008565:Membrane Proteins
keywords: Fluorescent dye; Melting temperature; Membrane protein; Protein stability; Thermostability assay