Isolation and western blotting of latex-bead phagosomes to track phagosome maturation

Anetta Härtlova, Julien Peltier, Orsolya Bilkei-Gorzo, Matthias Trost (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

2 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationPhagocytosis and phagosomes
Subtitle of host publicationmethods and protocols
EditorsRoberto Botelho
Place of PublicationNew York
PublisherSpringer
Pages241-248
Number of pages8
ISBN (Electronic)9781493965816
ISBN (Print)9781493965793
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in molecular biology
PublisherSpringer
Volume1519
ISSN (Print)1064-3745

Keywords

  • Phagosomes
  • Organelle purification
  • Phagocytosis
  • Western blotting

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