Minute™ Plant Endosome Enrichment Kit (20 preps)

 Manual & Protocol | MSDS

An understanding of endomembrane processes and their biological implications is important for understanding plant growth and development. Endosomes are a heterogeneous collection of organelles involved in the sorting and delivery of internalized material from the cell surface and the transport of materials from the Golgi to the lysosome or vacuole. In plant cells, two clearly defined endosomal compartments have been identified: the trans-Golgi network (early endosome equivalent) and the 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 amounts of starting material are usually required (10-50 g). We have developed this kit using a spin-column-based format coupled with selective precipitation for enrichment of the endosomal compartment. This approach is easy and rapid, and only a 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.

Kit includes: