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Isolation and Western Blotting of Latex-Bead Phagosomes to Track Phagosome Maturation

Lookup NU author(s): Dr Anetta Svitorka Hartlova, Dr Julien Peltier, Professor Matthias TrostORCiD

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Abstract

Phagocytosis plays an essential role in the immune system for the defense against invading microorganisms and the clearing of apoptotic cells. After internalization, the newly formed phagosome is constantly remodeled by fusion with early endosomes, late endosomes, and lysosomes. These changes ultimately deliver the engulfed material into the terminal degradative compartments known as phagolysosomes. However, defective phagosome maturation can result in inflammatory or autoimmune disease depending on the type of phagosome cargo. Therefore, characterization of the components involved in phagosome formation and maturation is important for a better understanding of macrophage physiological and pathological functions. In this chapter we describe a step-by-step protocol for the isolation of large-scale latex/polystyrene bead phagosome preparations with high degrees of purity for Western blotting analysis of phagosome maturation.


Publication metadata

Author(s): Härtlova A, Peltier J, Bilkei-Gorzo O, Trost M

Editor(s): Roberto Botelho

Publication type: Book Chapter

Publication status: Published

Book Title: Phagocytosis and Phagosomes: Methods and Protocols

Year: 2017

Volume: 1519

Pages: 241-248

Print publication date: 07/04/2017

Online publication date: 05/11/2016

Acceptance date: 17/08/2016

Publisher: Springer

Place Published: New York

URL: https://doi.org/10.1007/978-1-4939-6581-6_16

DOI: 10.1007/978-1-4939-6581-6_16

PubMed id: 27815884

Library holdings: Search Newcastle University Library for this item

ISBN: 9781493965816


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