Chapter 2 Digitonin Permeabilization Procedures for the Study of Endosome Acidification and Function
1989; Elsevier BV; Linguagem: Inglês
10.1016/s0091-679x(08)61600-3
ISSN0091-679X
Autores Tópico(s)Ion channel regulation and function
ResumoThis chapter describes a method for the study of endosome acidification using a digitonin-permeabilized cell assay. Digitonin can be used to permeabilize the plasma membrane of rabbit alveolar macrophages selectively without affecting intracellular membrane integrity. Endosomes remain functionally intact after digitonin permeabilization. This technique has proved to be an easy and rapid procedure to gain access to the cytoplasmic side of endosomes. Because other cell types are also amenable to selective digitonin permeabilization, this technique for plasma membrane permeabilization should be generally applicable. This technique can be applied to the study of other endosomal functions. Studies of anion and cation fluxes across the endosomal membrane should be possible. Specifically, the transport of Ca2+ could be addressed by this method because the mannose receptor exhibits Ca2+- dependent ligand binding.
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