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Eosinophils pp 93-100 | Cite as

Isolation and Functional Assessment of Eosinophil Crystalloid Granules

  • Renata Baptista-dos-Reis
  • Valdirene S. Muniz
  • Josiane S. NevesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1178)

Abstract

Cell-free granules, upon extrusion from human eosinophils, remain fully competent to secrete granule-derived proteins in receptor-mediated processes in response to different stimuli. However, in order to avoid the shrinkage and damage of granules, as well as preserve their structure, properties, and functionality, the use of an optimized process of subcellular fractionation using an isoosmotic density gradient is needed. Here, we describe a detailed protocol of subcellular fractionation of nitrogen-cavitated eosinophils on an isoosmotic iodinated density gradient (iodixanol) and the isolation of well-preserved and functional membrane-bound specific granules.

Key words

Granules Iodixanol Subcellular fractionation Isoosmotic gradient Eosinophils 

Notes

Acknowledgment

The authors thank Conselho Nacional de Pesquisas (CNPq—Brazil), Comissão de Aperfeiçoamento de Nível Superior (Capes—Brazil), and Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ—Brazil) for the financial support.

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Renata Baptista-dos-Reis
    • 1
  • Valdirene S. Muniz
    • 1
  • Josiane S. Neves
    • 2
    Email author
  1. 1.Institute of Biomedical SciencesFederal University of Rio de JaneiroRio de JaneiroBrazil
  2. 2.Institute of Biomedical SciencesFederal University of Rio de JaneiroRio de JaneiroBrazil

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