Stopped Flow Time‐Resolved Fluorescence Study of Serum Albumin–Curcuminoid Binding

Rapid mixing accessories to perform stopped flow measurements have found application in characterizing interactions and reactions occurring in solution. Reactants are expelled from syringes, mixed and injected into a flowcell. The flow is then stopped with the ensuing reaction / interaction monitored. If one of the reactants is fluorescent this phenomenon maybe used to follow the interaction. However, intensity based approaches can be influenced by fluctuations in the detected signal, eg. caused by photobleaching. This can be avoided by making use of the fluorescence lifetime, but data collection times have limited its usage. The recent introduction of very low deadtime electronics coupled with high repetition rate excitation sources have now allowed efficient and fast measurement of fluorescence decays. Here we monitor the interaction of a fluorescent curcuminoid mixture with serum albumin using a stopped flow accessory on a DeltaFlex system with DeltaDiode excitation at 100MHz.

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