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  • Biochemical isolation of insoluble tau in transgenic mouse models of tauopathies.

Biochemical isolation of insoluble tau in transgenic mouse models of tauopathies.

Methods in molecular biology (Clifton, N.J.) (2012-04-25)
Carl Julien, Alexis Bretteville, Emmanuel Planel
ABSTRACT

Tau is a highly soluble microtubule-associated protein (MAP) that is abundant in the central nervous system and expressed mainly in neuronal axons. Intracellular aggregates of insoluble tau protein are present in a group of neurodegenerative diseases called tauopathies, which include Alzheimer's disease. Numerous transgenic mouse models of tauopathies have been produced in the last decade, and analysis of insoluble tau in these animals has provided a powerful tool to understand the development of tau pathology. In this short chapter, we aim at reviewing the two main isolation methods, sarkosyl and formic acid extraction (and their variations), used for the biochemical isolation of insoluble tau in transgenic mouse models of tauopathy, and discuss their advantages and drawbacks.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Sodium orthovanadate, ≥90% (titration)
Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥98.5% (GC)
Sigma-Aldrich
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
N,N′-Methylenebis(acrylamide), 99%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, BioUltra, ≥99% (titration)