In Vitro Aggregation Assays Using Hyperphosphorylated Tau Protein

The overall goal of this procedure is to monitor and compare the aggregation kinetics of unmodified and hyper phosphorylated tau protein species. This is accomplished by first generating unmodified and hyper phosphorylated tau protein by the zippers assisted catalyst system. The second step is to set up the aggregation assays to be suitable for either of two different instruments.

The tau aggregation process is monitored by the fluorescence of two different thio. Flavin dies, theof flavin S and theof flavin T.The final step is to plot and compare the aggregation curves. Ultimately, the kinetic changes of thio flavin fluorescence from binding to aggregated TA species are used to show the hyper phosphorylation dependent enhancement of tau fibrillation in vitro.