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Updated: Aug 1, 2026

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
Published on: December 27, 2016
Protocol for the removal of bound endotoxin from tau protein produced in Escherichia coli
Andrew Shultz1, Daya Mena2, Jessica L Roy1
1Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, Amherst, MA, USA; Molecular and Cellular Biology Graduate Program, University of Massachusetts, Amherst, Amherst, MA, USA.
Abstract:
Tau is a widely studied protein that has implications for the progression of neurodegenerative disease. Here, we present a protocol for removing bound endotoxin from Escherichia coli (E. coli)-derived tau protein and verifying endotoxin removal. We describe steps for Triton X-114 treatment, seeding of HEK-Blue hTLR4 with endotoxin standards, collecting conditioned media, and endotoxin quantification. We then detail procedures for stimulating microglia and detecting cytokine secretion. This protocol is crucial for research studies that utilize tau in cell culture. For complete details on the use and execution of this protocol, please refer to Shultz et al.1.
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