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- W1580143657 abstract "Publisher Summary The fractionation and isolation of subcellular organelles have always been a major challenge in cell biology. Membrane-bound compartments are separated on the basis of their differential physical features, including shape, density, and charge. This chapter describes the procedures for the fractionation and isolation of Golgi and endosomal membranes along with the systematic methods for quickly optimizing sucrose-step gradients for separating marker proteins, as well as methods for quantitative immunoisolation of organelles with antibody-coated magnetic beads. In the case of the Golgi/endosomal system, the fractionation and coimmunoisolation of marker proteins can provide a comprehensive picture of the distribution of marker proteins in distinguishable types of membranes, yielding information about both compartmental identity and the mechanisms of localization of proteins to compartments. The colocalization of proteins can also be assessed at an ultrastructural level of resolution using immunogold electron microscopy. The chapter discusses the separation and distinction of Golgi and endosomal compartments using equilibrium density gradients. It also discusses the immunoaffinity isolation of organelles using magnetic beads and of immunoaffinity isolation of membranes and coimmunoisolation of marker proteins." @default.
- W1580143657 created "2016-06-24" @default.
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- W1580143657 date "2002-01-01" @default.
- W1580143657 modified "2023-09-24" @default.
- W1580143657 title "Separation of golgi and endosomal compartments" @default.
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- W1580143657 doi "https://doi.org/10.1016/s0076-6879(02)51857-7" @default.
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