PELs subcellular fractionaction

ganter ganter at interia.pl
Sun Sep 5 10:28:27 EST 2004


R.Jayakumar at RoswellPark.org ("Jayakumar, R") wrote in message news:<97101976F8A044468CA74FE11883B90E07496361 at VISTA.roswellpark.org>...
> I have done tens of subcellular fractionation of human cells.  So what 
> sort of cells are we looking at and what level of purity of fractions 
> are we talking of.  What sort of fractions do you want from the cells?  
> Are they human cells, are they suspension cell cultures or monolayers or 
> tissue samples? Try giving all the information you need before putting 
> up a question like that.  It should help to get help fast.
> Jai
>  
> > ----------
> > From: 	owner-methods at hgmp.mrc.ac.uk on behalf of ganter
> > Sent: 	September 3, 2004 11:44 AM
> > To: 	methods at hgmp.mrc.ac.uk
> > Subject: 	PELs subcellular fractionaction
> > 
> > Any useful protocol for subcellular fractionation of cells latently
> > infected by KSHV????
> > 


I wanted isolate separately cytoplasmic and nuclear proteins from
cells. These are human primary effusion lymphomas cells, which are
suspension cell cultures and latently infected by HHV8 (KSHV).The
specific problem is that my cytoplasmic extract is always contaminated
by proteins from nuclei fraction.(has been checked by cytosolic and
nuclear markers, by Western)
I tried many, many different options i.e  hypotonic buffers, various
detergent and many variants of salt concetration....

ganter



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