Each melting curve was baseline corrected and normalized to depict the fraction of bound strands on the y axis. The association constant, Ka, was quantified from the Arrhenius plot following the description in the literature,34 (link) using an in-house written LabVIEW routine (LabVIEW 2010, National Instruments). The dissociation rate, koff, was then quantified from Ka = kon/koff, where the association rate, kon (0.4 μM–1 s–1), was taken from the literature.26 (link),27 (link)
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Quantifying Biomolecular Binding Kinetics
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Corresponding Organization : München Klinik
Other organizations : Technical University of Munich, Max Planck Computing and Data Facility
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Variable analysis
independent variables
- Temperature
- Fraction of bound strands
- Association constant (Ka)
- Dissociation rate (koff)
- Baseline correction and normalization of melting curves
- Association rate (kon) from literature
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