Native protein denaturation using urea. (January 2017)

• Record Type: Journal Article
• Authors: Biggar, Kyle K.; Dawson, Neal J.; Storey, Kenneth B.
• Is Part Of: Biotechniques. Volume 62:Number 1(2017)
• Page Start: xiii
• Publication Date: 2017-01
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000114501
• Languages: English
• ISSNs: 0736-6205

Abstract

Protocol Summary: Here we present a new protocol to analyze protein unfolding kinetics using a quantified real-time thermocycler. This technique enables the analysis of a wide range of denaturants (and their interactions with temperature change) on protein stability in a multi-well platform, where samples can be run in parallel under virtually identical conditions and with highly sensitive detection. Using this set-up, researchers can evaluate the half-maximal rate of protein denaturation (Knd), maximum rate of denaturation (Dmax), and the cooperativity of individual denaturants in protein unfolding (µ-coefficient).