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).

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Journal BioTechniques
Biggar, K.K, Dawson, N.J. (Neal J.), & Storey, K. (2017). Native protein denaturation using urea. BioTechniques, 62(1). doi:10.2144/000114501