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

Lu Falong via proteins%40net.bio.net (by FLLu At genetics.ac.cn)
Sun Nov 26 02:00:34 EST 2006


proteins-request,

	I have met a similar problem. Membrane I used was 0.45um pore size PVDF from Pierce. After transfer, the prestained ladder could be seen, but after a brief rinse, most of the prestained ladder was gone (Transfer buffer Towbins buffer with 10% methanol and 0.05% SDS). Also I have never seen such 
things with NC membrane. I am wondering if it is caused by the SDS present in the buffer. However, in the instruction of PVDF membrane from biorad, methanol and SDS would not interfere binding of proteins to PVDF membrane. Any comments or suggestions?

==== 2006-11-26 01:01:31 ======

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>   1. why I can see my band dyed in proceau S on PDFC?
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>----------------------------------------------------------------------
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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
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>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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Sincerely,¡¡¡¡¡¡¡¡¡¡

Lu Falong
 Group 808
 Institute of Genetics and Developmental Biology, CAS 
 Beijing, PR China
 FLLu At genetics.ac.cn
 2006-11-26  




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