Minute¢â Endosome Isolation and Cell Fractionation Kit
Early endosomes (EE) provide the starting point for late endosome maturation. The EEs are mainly derived
from primary endocytic vesicles that fuse with each other. EEs receive endocytic cargo not only through the
clathrin-mediated pathway but several other pathways. In addition to their roles in normal cell physiology,
endocytic processes play a key role in many diseases such as Alzheimerâ¢æ¢âs disease and inherited
lysosomal storage diseases. Traditional methods for isolating endosomesare based on density gradient
ultracentrifugation. The protocol requires large amount of starting material and the methods are tedious and
time consuming. The minute Endosome Isolation and Cell Fractionation Kit provides a spin-column based
novel endosome isolation technology that is rapid, simple and requires smaller number of cultured cells or
milligram amounts of tissues. This kit can precipitate and significantly enrich early endosomes from cultured
cells or tissues. The availability of the kit should facilitatethe research in the field.
Enrichment of endosomes. Endosomes were isolated from mouse kidney tissue as described in the
protocol. Total cellular protein(T) and endosomes(E) in duplicate were separated in 12% SDS-PAGE. After
electrical trasnfer, the membrane was stained with Ponceau Red(A). The blot was probed with early
endosome antigen1, Na/K ATPase and tubulin by Western blotting(B).
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