Since your technique works in for NC then obviously something is wrong
with your PVDF. Have you hydrated your PVDF in methanol before transfer?
This might a source of the problem
-----Original Message-----
From: proteins-bounces At oat.bio.indiana.edu
[mailto:proteins-bounces At oat.bio.indiana.edu] On Behalf Of
proteins-request At oat.bio.indiana.edu
Sent: Saturday, November 25, 2006 7:01 PM
To: proteins At magpie.bio.indiana.edu
Subject: Proteins Digest, Vol 18, Issue 13
Send Proteins mailing list submissions to
proteins At net.bio.net
To subscribe or unsubscribe via the World Wide Web, visit
http://www.bio.net/biomail/listinfo/proteins
or, via email, send a message with subject or body 'help' to
proteins-request At net.bio.net
You can reach the person managing the list at
proteins-owner At net.bio.net
When replying, please edit your Subject line so it is more specific
than "Re: Contents of Proteins digest..."
Today's Topics:
1. why I can see my band dyed in proceau S on PDFC?
(=?GB2312?B?veDV1A==?=)
----------------------------------------------------------------------
Message: 1
Date: Sat, 25 Nov 2006 10:57:51 +0800
From: "=?GB2312?B?veDV1A==?=" <janejiesweet At gmail.com>
Subject: [Protein-analysis] why I can see my band dyed in proceau S on
PDFC?
To: proteins At magpie.bio.indiana.edu
Message-ID:
<4d0022c90611241857v48f5c87dj34e2c08f2df449ac At mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
sir/madam:
I have been trying to blot proteins of brain from an 8% SDS PAGE
toBioRad PVDF membrane. I thought that my blot was unsuccessful
afterattempting to visualize by Ponceau S. I could not even detect my
MWstandards.but the same process worked well in NCmembrane,where the
band
showed clearly.. Any clues as to why my ponceau S did not work? I
diluted a
fresh Sigam concentrate (20 mL) into 180 mL ofdeionized water. I stained
for
5 min and watch as the membrane destained in 1% AcOH. At no time did I
see
anything resembling a protein band. This problem has occured for the
past
two consecutive times that I have tried the technique. Your comments are
appreciated.
------------------------------
_______________________________________________
Proteins mailing list
Proteins At net.bio.nethttp://www.bio.net/biomail/listinfo/proteins
End of Proteins Digest, Vol 18, Issue 13
****************************************
