Stopped-flow Kinetics

Idea: In this lab course we investigate the unfolding and refolding kinetics of lysozyme
using a stopped-flow apparatus

Biophysics Lab Course, Institute of Biophysics, Ulm University


Unfolding and refolding kinetics of lysozyme is studied by rapid mixing of a protein solution with a buffer solution of different denaturant concentration.

The change of intrinsic tryptophane fluorescence is monitored to follow the changing fractions of folded and unfolded molecules.

Rate coefficients for unfolding and refolding are determined from a 'chevron plot' of the apparent rate coefficients, assuming a two-state model for protein folding.