protein and RNA purification

Stephen R. Lasky, Ph.D. Stephen_Lasky at brown.edu
Thu Dec 22 09:16:01 EST 1994


In article <3daac4$1tu at neuro.usc.edu>, william at neuro.usc.edu (Fiberman) wrote:

> Is there a protocol to get proteins and RNA from the same tissue in one  
> step?  If you know of one, or can provide a reference, please let me know!
> Thanks.
> 
> -fm

I don't know of one that gives you proteins and RNA in one step but there
are several that give you both from the same cells at the same time in
several steps:

Trizol (many of the chomzinski (sic) kits) allow you to isolate RNA, DNA,
and protein in reasonable lengths of time.  I'm sure that the proteins are
inactive but they can be used for Westerns.

I isolate nuclear proteins (transcrption factors) using a modified Dignam
tech and then use the cytosolic fraction for and RNA prep by extracting
with phenol/chloroform/isoamylalcohol.  

You can get nuclear proteins and RNA as above but from smaller numbers of
cells using NP-40 lysis techniques, I don't have a ref for this.  We have
found that the modified Dignam and NP-40 lysis techniques extract nuclear
proteins with different efficiencies.

I guess a lot depends upon what kind of proteins you want.

Hope that helps.

SRLasky

-- 
*********************************************************************
Stephen R. Lasky, Ph.D.
Roger Williams Medical Center/Brown University
Phone: 401-456-6572       Fax: 401-456-6569       e-mail: Stephen_Lasky at brown.edu
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
"To me at least, 'Yuck' doesn't capture the full essence of death by
neurotoxin."   -Dick Dunn
*********************************************************************



More information about the Methods mailing list