Lamberti 2015 Cold Spring Harb Protoc: Difference between revisions
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|year=2015 | |year=2015 | ||
|journal=Cold Spring Harb Protoc | |journal=Cold Spring Harb Protoc | ||
|abstract=Immortalized macrophage lines and primary macrophages display the ability to internalize small latex beads through the endocytic pathway. This protocol describes a simple and robust method for separating endocytic organelles from macrophages on a sucrose gradient, taking advantage of the significantly lower density of the organelles containing latex beads compared with other intracellular organelles. The latex beads are retained in the endosomes as they mature; therefore, harvesting cells at different time points after internalization permits the purification of different organelle fractions, particularly early and late endosomes | |abstract=Immortalized macrophage lines and primary macrophages display the ability to internalize small latex beads through the endocytic pathway. This protocol describes a simple and robust method for separating endocytic organelles from macrophages on a sucrose gradient, taking advantage of the significantly lower density of the organelles containing latex beads compared with other intracellular organelles. The latex beads are retained in the endosomes as they mature; therefore, harvesting cells at different time points after internalization permits the purification of different organelle fractions, particularly early and late endosomes. | ||
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{{Labeling | {{Labeling | ||
|organism=Mouse | |organism=Mouse | ||
| | |tissues=Macrophage-derived | ||
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Latest revision as of 16:10, 9 November 2016
Lamberti G, de AraΓΊjo ME, Huber LA (2015) Isolation of macrophage early and late endosomes by latex bead internalization and density gradient centrifugation. Cold Spring Harb Protoc 2015(12):pdb.prot083451. |
Lamberti G, de Araujo ME, Huber LA (2015) Cold Spring Harb Protoc
Abstract: Immortalized macrophage lines and primary macrophages display the ability to internalize small latex beads through the endocytic pathway. This protocol describes a simple and robust method for separating endocytic organelles from macrophages on a sucrose gradient, taking advantage of the significantly lower density of the organelles containing latex beads compared with other intracellular organelles. The latex beads are retained in the endosomes as they mature; therefore, harvesting cells at different time points after internalization permits the purification of different organelle fractions, particularly early and late endosomes.
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Organism: Mouse
Tissue;cell: Macrophage-derived