[Protein-analysis] RE: why I can see my band dyed in proceau S on PDFC?

Tsalavos Sotiris via proteins%40net.bio.net (by tsalavos At fleming.gr)
Mon Nov 27 10:46:35 EST 2006



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

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Subject: Proteins Digest, Vol 18, Issue 13

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Today's Topics:

   1. why I can see my band dyed in proceau S on PDFC?
      (=?GB2312?B?veDV1A==?=)


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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.


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