Volume 5, Number 1, pp. 9-12
Isolation of raft microdomains from rat brain: a novel fast method based on distribution in a dextran-polyethylenglycol biphasic system
Yu.Ya. Kit1, V.A. Kovalyova2, A.A. Kim3 and R.S. Stoika1
1
Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, Ukraine
2
Ukrainian State University of Forestry and Wood Technology, Lviv, Ukraine
3
Institute of Nuclear Physics, Academy of Sciences of Uzbekistan, Ulugbek, Tashkent, Uzbekistan
Raft microdomains are detergent-resistant membrane fragments of eukaryotic cells, which contain cholesterol, sphingolipids and proteins. They are dynamic structures involved in the regulation of cellular activity. Usually, the method of flotation analysis is applied for the isolation and study of raft microdomains. It is based on a lengthy isopycnic centrifugation. We developed a novel, convenient and rapid method for the isolation of raft microdomains from rat brains, based on the distribution of detergent-soluble and detergent-insoluble membrane components in the biphasic dextran-polyethylenglycol system. Thin-layer chromatography of the lipids and two-dimensional electrophoresis of the proteins have shown the presence of cholesterol and protein complexes in the isolated rat brain raft microdomains.
Keywords: biphasic system, brain, raft microdomains, rat, isolation