Removal of proteins from membranes->references

[Bradley Turner]

Dear Johannes,

Yes, it is possible to recover proteins blotted to either PVDF or NC.
I do it quite often to obtain small amounts of highly purified proteins.
I've listed some references below. 

In general, one seems to get higher protein recoveries
from NC but often they are slightly contaminated with small amounts of
dissolved NC-one gets less recovery from PVDF but there is far less
contamination from the membrane.


REFERENCES:

The recovery of nitrocellulose-bound proteins
Analytical Biochemistry 148:105-110, 1985
PJ Anderson

Preparative elution of proteins from nitrocellulose membranes after
separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis
Analytical Biochemistry 148:87-92, 1985
BS Parekh, HB Mehta, MD West, RC Montelaro

Preparative elution of proteins blotted to Immobilon membranes 
Analytical Biochemistry 168:48-53, 1988
B Szewczyk, D Summers

Protein antigen purification by preparative protein blotting
Electrophoresis 8:432-438, 1987
RC Montelaro

Methods for the isolation of proteins immobilized on S&S NC nitrocellulose
Sequences: application update number 388
Schleicher and Schuell, Inc
Keene, NH 03431 USA 800-245-4024
D-3354 Dassel, West Germany
http://www.s-and-s.com

Quantitative recovery of proteins from Immobilon-P blots
N Legendre
Application note RP245
Millipore Corporation Bedford MA 01730 617-645-5476
http://www.millipore.com

On the electrotransfer of polypeptides from gels to nitrocellulose
membranes 
Analytical Biochemistry 128:302-311, 1983
W Lin, H Kasamatsu

Proteins transferred to nitrocellulose for use as immunogens
KA Knudsen
Analytical Biochemistry 147,:285-288, 1985

Use of blotted proteins as immunogens
B Szewczyk, DR Harper
Chapter 15, pages 189-205 in
Protein Blotting: A practical approach
editor Bonnie S Dunbar
IRL Press/Oxford University Press

Specialized aspects of protein blotting
ER Tovey, BA Baldo
Chapter 3, pages 43-68
in Protein Blotting: methodology, research, and diagnostic applications
editors: BA Baldo, ER Tovey
Karger press

I hope this helps,
Brad Turner


**************************************************************
		    Bradley Turner
	        Beth Israel Deaconess
                    Medical Center

Harvard Medical School		617-667-1215 phone
Division of Gastroenterology	617-667-2767 fax
Room Dana 536			bsturner at mbcrr.harvard.edu
330 Brookline Avenue		bturner at bidmc.harvard.edu
Boston, MA 02215, USA		turner at sprcore.bidmc.harvard.edu
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>From johannes at biobase.dk Thu Apr 17 10:36:55 1997
To: methods at net.bio.net
From: Johannes Dalgaard Clausen <johannes at biobase.dk>
Subject: Removal of proteins from membranes
Date: Thu, 17 Apr 1997 16:21:21 +0100
NNTP-Posting-Host: majestetix.medmicro.aau.dk
Mime-Version: 1.0
Content-Transfer-Encoding: 7bit

Hi,

Does anyone know if it is possible to remove proteins from a membrane
(nitrocellulose or PVDF) after the proteins have been transferred to the
membrane? I have tried boiling the membrane-fragments for 5 min in a SDS
sample buffer (125 mM Tris-HCl pH 6.8, 10% glycerol, 2.3% SDS, 5%
beta-mercaptoethanol), but this didn't seem to have any effect.

If you have any experience with this problem, please reply to
johannes at biobase.dk.


Johannes D. Clausen
Dept. of Medical Microbiology and Immunology
University of Aarhus, Denmark



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