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Anal Biochem. 1993 Oct;214(1):205-11.

Isolation of the phagocytic compartment from macrophages using a paramagnetic, particulate ligand.

Author information

1
Department of Ophthalmology and Visual Sciences, Kentucky Lions Eye Research Institute, University of Louisville School of Medicine 40292.

Abstract

Isolation of Fc receptor-dependent phagosomes from a macrophage cell line, J774 A.1, was accomplished using antibody-opsonized, paramagnetic beads. Following binding and ingestion of these beads, cells were homogenized in a standard membrane isolation buffer. Phagosomes containing the trapped paramagnetic beads were isolated by subjecting the whole cell homogenate to a magnetic field. The method is extremely simple and the preparation of an enriched phagosome fraction from a whole cell homogenate is rapid and highly selective. The method should provide useful starting material for investigators interested in cytoskeletal involvement in phagocytosis, in kinetic studies of ingestion, in phagosome-lysosome fusion, and in the ability of a particular ligand to initiate phagocytosis.

PMID:
8250224
DOI:
10.1006/abio.1993.1478
[Indexed for MEDLINE]

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