Methods Digest, Vol 49, Issue 12

[TieQiao Wu]

Hi, 

Did you use prestained protein marker? You should see the marker on your
membrane very clearly using your transferring voltage and time setting.
What I can suggest at the moment is that you check your transfer buffer
and your sample buffer.

Cheers   

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Today's Topics:

   1. Western Blotting problems (Alsagaby, Suliman)


----------------------------------------------------------------------

Message: 1
Date: Sat, 13 Jun 2009 19:16:37 +0100
From: "Alsagaby, Suliman" <salsag from essex.ac.uk>
Subject: Western Blotting problems
To: <methods from magpie.bio.indiana.edu>
Message-ID:
	<EC159B4211898E43ADF0C4F5C82567EE03DA5446 from sernt4.essex.ac.uk>
Content-Type: text/plain;	charset="us-ascii"

Hello

I did western blot twice and every time I have the same problem in which
the PVDF membrane does not contain any protein although it contains
protein marker. The bands were visualized by staining the gel. So it
seems to me that the bands did not transfer to the membrane. I verified
the transfer duration: one for 12h at 30V and the other one was for 1h
at 100V.

I am great full if any body can help me and direct me to the right
protocol if I am doing wrong one. 



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