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An understanding of endomembrane processes and their biological implication is important for understanding

plant growth and development. Endosomes are a heterogeneous collection of organelles in the sorting and

delivery of internalized material from cell surface and the transport of materials from Golgi to lysosome or

vacuole. In plant cells, two clearly defined endosomal compartments have been identified: the trans-Golgi

network (early endosome equivalent) andthe multivesicular body (late endosome equivalent). Traditionally,

methods for isolation and purification of these two compartments are tedious and time-consuming involving

sucrose gradient centrifugation. In some cases, sucrose gradient isolation is coupled with immunoaffinity

purification and larger amount of starting materials are usually required (10-50 g). We have developed this kit

using a spin-column-based format coupling with selective precipitation for enrichment of endosomal

compartment. This approachis easy and rapid and only small amount of starting material is required.

Detection of Enriched Endosomes from N. benthmiana by  Western blotting. The plant leaf that carries GFP-

tagged  ara6 (early endosome marker) was processed as described in the protocol and probed with anti-

ara6 and anti-histone  antibodies. Lane 1, Total tissue lysate; lane 2, Supernatant  from step 4 of the protocol;

lane 3, Enriched endosome fraction.