Protocol for expressing and purifying recombinant full-length Tau by combining affinity chromatography with preparative HPLC

Deposits of microtubule-associated protein full-length Tau (Tau2N4R) are implicated as a hallmark of Alzheimer's disease. Its biochemical and structural characterization is key to understanding disease progression, aggregate toxicity, and designing therapeutics. We present a protocol for expressing and purifying Tau2N4R by combining affinity chromatography with preparative high-performance liquid chromatography (HPLC). We describe steps for linking an N-terminal hexahistidine tag and a cleavable SUMO tag to Tau2N4R. This purification ensures an ultrapure Tau2N4R that gives rise to heparin-induced and cofactor-free in vitro fibrillation.