Related Experiment Video
Updated: Jun 18, 2026
12:47
Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
Published on: December 27, 2016
Protocol for expressing and purifying recombinant full-length Tau by combining affinity chromatography with
Ruiheng Jing1, Sayanta Mahapatra1, Abhishek Bimali1
1University of Colorado, Department of Chemistry, Boulder, CO 80309, USA.
STAR Protocols
|June 16, 2026
Summary
Researchers developed a new method to purify Tau2N4R, a protein linked to Alzheimer's disease. This ultrapure Tau protein is crucial for studying disease mechanisms and developing new treatments.
Area of Science:
- Neuroscience
- Biochemistry
- Molecular Biology
Background:
- Microtubule-associated protein Tau (Tau2N4R) deposits are a hallmark of Alzheimer's disease (AD).
- Understanding Tau's biochemical and structural properties is vital for AD research, including aggregate toxicity and therapeutic development.
Purpose of the Study:
- To present a robust protocol for the expression and purification of full-length Tau2N4R.
- To obtain ultrapure Tau2N4R for in vitro studies of fibrillation.
Main Methods:
- The protocol involves expressing Tau2N4R with an N-terminal hexahistidine tag and a cleavable SUMO tag.
- Purification is achieved through a combination of affinity chromatography and preparative high-performance liquid chromatography (HPLC).
Main Results:
- The described method yields ultrapure Tau2N4R.
- The purified Tau2N4R protein undergoes heparin-induced and cofactor-free in vitro fibrillation.
Conclusions:
- This protocol provides a reliable method for obtaining high-purity Tau2N4R.
- The availability of pure Tau2N4R facilitates further investigation into Alzheimer's disease pathogenesis and therapeutic strategies.
