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Magnetic Isolation of Phagosomes Containing Toll-Like Receptor Ligands

  • Tanja Petnicki-Ocwieja
  • Linden T. Hu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1690)

Abstract

Phagosomal compartments are critical in microbial defense as vesicles that degrade invading organisms. In a broader context, vesicular trafficking plays an important role in shuttling many different types of cargo that are critical for proper function of the cell. Endosomal and phagosomal vesicles are thus important locations for the assembly of intracellular signaling platforms that mediate host responses to invasive pathogens such as Borrelia burgdorferi. Isolation of phagosomes from cells is an important technique that allows for a detailed study of phagosomal components and signaling complex assembly. However, purification of phagosomes had previously been challenging and it has been difficult to obtain sufficient purity of the phagosomal fractions. Here, we modify a new magnetic isolation technique that greatly simplifies purification of phagosomes and isolates vesicles with sufficient purity for analysis.

Key words

Phagocytosis Phagosomes Magnetic beads Toll-like receptors TLR2 Lyme disease Borrelia burgdorferi 

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.Department of Molecular Biology and MicrobiologyTufts University School of MedicineBostonUSA

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