Lipid Rafts and Immunoblotting

dowlinj3 from gmail.com via methods%40net.bio.net (by dowlinj3 from gmail.com)
Wed Jun 20 13:07:17 EST 2007


I am currently working on some lipid raft experiments by sucrose
geadients with 1% v/v T-X100. I would like to probe for markers such
as GM-1 ad Flotillin. However, when I precipitate protein from the
fractions by TCA or concentrate them by vivaspin I get insufficinet
protein (Comassie staining) on my SDS-PAGE gels to go on to do
westerns. Is there a better way to precipitate the proteins. I find
with the TCA its not great to have to resuspend your samples in SDS
Loading buffer as you can't do a BCA assay or use them again!
Any help/comments are aprreciated - Thanks
Jen



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