problem

Peter pxpst2 at spam.suxs.unixs.cis.pitt.edu
Sun Jul 19 21:36:59 EST 1998


In article <pxpst2-1907982128150001 at pelli.pathology.pitt.edu>,
pxpst2 at spam.suxs.unixs.cis.pitt.edu wrote:

> In article <g.s.stewart-1907981751050001 at med179.bham.ac.uk>,
> g.s.stewart at bham.ac.uk (Grant Stewart) wrote:
> 
> > When I am preparing samples for Western I do the following:
> > 
> > Thaw pellet of gamma-radiation treated LCLs (~4,000,000 cells) at 37oC.
> > Resuspend pellet in 200ul UTB buffer (9M urea, 150mM B-mercaptoethanol,
> > 50mM Tris pH7.5). Sonicate for 2x15 seconds, keeping cool, then centrifuge
> > for 25mins at 35000 rpm at 10oC. 
> > 
> > The problem is that after centrifugation I get this viscous bubbly layer
> > floating on the top of the centrifuged lysate. There is a pellet therefore
> > it has been centrifuged properly. 
> 
> 
> What about Lipids?


think about how soap is made and you may see were your problems are.

;-)


peter

-- 
"Don't you eat that yellow snow
            watch out where the Huskies go"    FZ

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