stripping Western blots

Scott Selby Med Heme selby at lenti.med.umn.edu
Wed Aug 10 16:52:07 EST 1994


FJ van der Wal (vdwal at bio.vu.nl) wrote:
: In article <94222.093716MSH4 at psuvm.psu.edu>, Peggy Halleck
: <MSH4 at psuvm.psu.edu> wrote:

: > Hi!  I'm looking for a reliable method to strip Western blots for reprobing.
: > I'm using Immobilon membranes and ECL detection with HRP-protein A.

: Successful and repeated stripping can be done according to the protocol in
: 'ECL Highlights' V4 p12 by Driver, Lee, Guilford and Price, (1992). We do
: it all the time!

: Stripping buffer: 100mM 2-mercaptoethanol, 2% SDS, 62.5mM Tris-HCl pH6.7

{snip}  Keep in mind, however, that 2-mercaptoethanol can alter your Ab 
binding site.  While this method works for some proteins, it may not work 
for yours.  I've had both experiences, good and bad, with this method.

Scott Selby
University of MN
Dept. of Med/Heme
Mpls., MN



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