Proteins from phenol

[Ed Rybicki]

Message-ID: <MAILQUEUE-101.921211170205.91 at micro.uct.ac.za>
To:           vioque at cica.es (Agustin Vioque),
              methods-reagents at net.bio.net
From:         "Ed Rybicki"  <ED at micro.uct.ac.za>
Date:         11 Dec 92 17:02:05 SAST-2
Subject:      recovery of proteins from phenol
Reply-to:     ed at micro.uct.ac.za
Priority:     normal
X-mailer:     Pegasus Mail v2.3 (R4).

> I have a highly purified protein that I want to silver stain on a gel. The
> protein is contaminated with a large amount of RNA which will also silver
> stain. Toget rid of the RNA I have tried a treatment with NaOH (10', 80C,
> 0.1 N) but a get a smear of partially degraded RNA upon staining of the gel.
> I am thinking of doing a phenol extraction, but how can I recover the protein
> from the phenol phase in a way suitable for SDS-PAGE? If anybody has an
> idea I would appreciate it very much. Sorry for my poor english.

Never apologise!  Let them read it and suffer, it makes them broader
minded!

But about phenol-solubilised proteins: easiest way to recover them is to
dilute with 5 vol of cold methanol, and spin: wash ppte with methanol, and
air-dry, and solubilise in SDS-PAGE sample buffer, with heating.  Works
better than acetone ppte. in my hands.  Hope this is of use,

  ____________________________________________________________________
 | Ed Rybicki, PhD             |     "Emancipate yourselves from      |
 | (ed at micro.uct.ac.za)        |            Mental Slavery            |
 | Dept Microbiology           |          None but ourselves          |
 | University of Cape Town     |         Can free our minds...."      |
 | Private Bag, Rondebosch     |                                      |
 | 7700, South Africa          |             - Bob Marley             |
 | fax: 27-21-650 4023         |          (Redemption Songs)          |
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  ____________________________________________________________________
 | Ed Rybicki, PhD             |     "Emancipate yourselves from      |
 | (ed at micro.uct.ac.za)        |            Mental Slavery            |
 | Dept Microbiology           |          None but ourselves          |
 | University of Cape Town     |         Can free our minds...."      |
 | Private Bag, Rondebosch     |                                      |
 | 7700, South Africa          |             - Bob Marley             |
 | fax: 27-21-650 4023         |          (Redemption Songs)          |
  --------------------------------------------------------------------