Dan,
I think you would have to prefractionate before digestion due to the protease
inhibitors present in the samples.
Mike Knierman
To: Recipients of ABRF List <abrf@aecom.yu.edu>
cc:
Subject: RE: Solution Digests
I haven't done it, but it seems to me that if you precipitate the proteins
in plasma or CSF with TCA or acetone, wash the precipitate, add the
appropriate buffer for the enzyme you want to use, neutralize solution if
too acidic and then follow a regular digest protocol, or if you want to
isolate a particular protein, solubilize the precipitate and separate the
different proteins by RP HPLC using a C4 or C8 column.
Amina
Amina S. Woods, Ph.D.
NIDA Intramural Program, NIH
5500 Nathan Schock Drive
Baltimore, MD 21224
Tel: 410-550-1507
Fax: 410-550-2971
e-mail: awoods@intra.nida.nih.gov <mailto:awoods@intra.nida.nih.gov>
-----Original Message-----
From: Association of Biomolecular Resource Facilities
[mailto:abrf-request@aecom.yu.edu]On Behalf Of Dan Kirby
Sent: Thursday, January 27, 2000 7:04 PM
To: Recipients of ABRF List
Subject: Solution Digests
I've scoured the ABRF archives as well as many of the Protocol Sites on the
Internet, but I can't find a protocol for solution digestion of proteins in
CSF and/or plasma. Can anyone provide either a protocol or a reference
thereto? Thanks in advance. Dan Kirby
_______________________
Dan Kirby
Praecis Pharmaceuticals
1 Hampshire Street
Cambridge, MA 02139
dan.kirby@praecis.com
<http://www.praecis.com>
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