I don’t have any information about affinity constants of the interaction I want to analyze. How should I design my kinetic experiment?
The switchBUILD kinetic tool helps to easily set up an optimized taskflow to analyze the interaction of interest and automatically suggests concentration ranges and measurement times based on the estimated affinity of the interaction partners.
When no affinity constants are known, an educated guess of the affinity range helps to set up your assay. This can be done, for instance by choosing an interaction with a comparable molecule class (e.g. antibody) from the provided drop-down list. Based on this estimate, an affinity scouting can be carried out. To do so, choose a high dilution factor (at least 3x) and at least three concentration steps to cover a possibly wide concentration range. This way, it is usually possible to roughly determine the affinity range which can be narrowed down in further experiments.