Abstract:

[Abstract]  Objective   To verify the validity of the sucrose density gradient ultracentrifugation method for lipid rafts
from cerebral cortex.Methods  Extract lipid rafts from cerebral cortex in mouse were extracted by the sucrose density gradient ultracentrifugation method. The properties of lipid rafts were detected by Western blotting method, double enzyme and light scattering methods. HPLC MS/MS proteomics and bioinformatics were used to locate proteins of lipid rafts in cells. Results Lipid rafts from cerebral cortex were provided with the model properties of lipid rafts such as high light scattering and cholesterol and high expression of Flotillin-1. HPLC MS/MS proteomics identified total 647proteins. Most of these proteins were from plasma membrane, endoplasmic reticulum, cytoskeleton and cytosol, however, there were 21% proteins among total647proteins were from nucleus, mitochondria and ribosomes.Conclusion  The sucrose density gradient ultracentrifugation method is a effective method to extract lipid rafts from cerebral cortex, however, the properties of mixture should be considered.

Key words: membrane lipids, computational biology, ultracentrifugation, hypothalamus, proteins, genome, lipid raft