Nuclear and cytoplasmic cell extracts
johnson_lab at qm.yale.edu
Tue Jun 6 23:17:23 EST 1995
In article <3r1qtb$1hi at rc1.vub.ac.be>, pberr at dec5 (Pascale Berr) wrote:
> I'm looking for a method to separate nuclear and cytoplasmic proteins
> from mammalian cells extracts.
> Any suggestion welcome!
For a "quick-and-(not so)-dirty", you could modify a popular nuclear
extraction protocol (Schreiber et al, Nucleic Acids Research 17:6419).
This is easy, gives good nuclear extracts for gel-shifts (DNA binding
proteins) and the cytoplasmic extract has RNA (as the authors point out)
and also yields a lot of protein. In western blotting for nuclear
proteins, I found plenty in the nuclear extract and only trace amounts in
the cytoplasmic extracts (and vice versa) taking no particular care to
rigorously separate the fractions.
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