How to isolate mammalian cell cytoplasm?

Wolfgang Schechinger wgschech at med.uni-tuebingen.de
Mon Nov 10 13:55:41 EST 1997


Hi. Buffers with 0.5 to 1% Triton -X-100 or Nonidet-P40 should work. 
If you treat your dishes for 10 min at 4degC, the nuclei will remain 
attached to the ground. If you have suspended cells, centrifuge the 
nuclei down and use the supernatant.
This procedure will dissolve the membranes, too.

Wolfgang

> Hello, can anyone help me?
> I need a protocol to isolate cytoplasmic extracts from mammalian
> culture cells. It is of particular importance that I minimise
> nuclear contamination. Thanks, RSVP mladomery at hgu.mrc.ac.uk
> 
> 
> 
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Wolfgang Schechinger         
University of Tuebingen
email: wgschech at med.uni-tuebingen.de
http://www.medizin.uni-tuebingen.de/~wgschech/research.htm
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