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Updated: Aug 13, 2026

The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking
Published on: February 12, 2022
Reconstitution of transport to recycling endosomes in vitro
René Bartz1, Corinne Benzing, Oliver Ullrich
1Department of Cell Biology, University of Texas, Southwestern Medical Center, Dallas, USA.
Abstract:
Using an in vitro assay instead of an approach in vivo greatly facilitates the analysis of complex intracellular mechanisms. This becomes particularly important for studying vesicular trafficking in both the endocytic and exocytic pathways with multiple transport routes connecting cellular organelles. Our chapter describes a novel cell-free assay that reconstitutes endosomal transport to recycling endosomes. The method measures transport of transferrin, a marker for endocytosis/recycling, from an endosome-enriched donor fraction to immunoisolated Rab11-positive acceptor recycling endosomes. Transfer of acridinium-labeled transferrin is detected by a highly sensitive chemiluminescence reaction using a luminometer.
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