Difference between revisions of "De Araujo 2015 Cold Spring Harb Protoc-A"

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de Araújo ME, Lamberti G, Huber LA (2015) Isolation of Early and Late Endosomes by Density Gradient Centrifugation. Cold Spring Harb Protoc 2015(11):pdb.prot083444.

» http://www.ncbi.nlm.nih.gov/pubmed/26527762

de Araújo ME, Lamberti G, Huber LA (2015) Cold Spring Harb Protoc

Abstract: Density gradient centrifugation is a common method for separating intracellular organelles. During centrifugation, organelles float or sediment until they reach their isopycnic position within the gradient. The density of an organelle depends on its content, size, shape, and the lipid:protein ratio. The degree of separation between different organelles will therefore be highly dependent on how different their isopycnic points are in a given buffer. Separation will also depend on the medium used to prepare the gradient, whether it is sucrose (the most common) or an alternative. Here we describe the use of both continuous and discontinuous (step) gradients to isolate endocytic organelles.

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 {{Publication
-|title=de Araújo ME, Lamberti G, Huber LA (2015) Homogenization of Mammalian Cells. Cold Spring Harb Protoc 2015(11):pdb.prot083436.
+|title=de Araújo ME, Lamberti G, Huber LA (2015) Isolation of Early and Late Endosomes by Density Gradient Centrifugation. Cold Spring Harb Protoc 2015(11):pdb.prot083444.
-|info=http://www.ncbi.nlm.nih.gov/pubmed/26527761
+|info=http://www.ncbi.nlm.nih.gov/pubmed/26527762
 |authors=de Araújo ME, Lamberti G, Huber LA
 |year=2015
 |journal=Cold Spring Harb Protoc
-|abstract=Homogenization is the name given to the methodological steps necessary for releasing organelles and other cellular constituents as a free suspension of intact individual components. Most homogenization procedures used for mammalian cells (e.g., cavitation pump and Dounce homogenizer) rely on mechanical force to break the plasma membrane and may be supplemented with osmotic or temperature alterations to facilitate membrane disruption. In this protocol, we describe a syringe-based homogenization method that does not require specialized equipment, is easy to handle, and gives reproducible results. The method may be adapted for cells that require hypotonic shock before homogenization. We routinely use it as part of our workflow to isolate endocytic organelles from mammalian cells
+|abstract=Density gradient centrifugation is a common method for separating intracellular organelles. During centrifugation, organelles float or sediment until they reach their isopycnic position within the gradient. The density of an organelle depends on its content, size, shape, and the lipid:protein ratio. The degree of separation between different organelles will therefore be highly dependent on how different their isopycnic points are in a given buffer. Separation will also depend on the medium used to prepare the gradient, whether it is sucrose (the most common) or an alternative. Here we describe the use of both continuous and discontinuous (step) gradients to isolate endocytic organelles.
 }}
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