Posted 31 August 2012 - 08:21 AM
I am using home-made 12% PAGEs, running at 100V (the buffer got really hot, could that be the problem?) for over an hour. I transfer overnight at 4oC onto nitrocellulose membrane.
My running buffer is pH8.3, and my transfer buffer (containing 10% MeOH) is pH8.38.
In the past, I have not run my PAGEs as long as I did recently, and think that could be the reason why (because the running buffer heats up).
Any suggestions or clues as to why this happened and how to avoid it would be greatly appreciated.
Thanks to everyone!
Posted 31 August 2012 - 09:30 AM
The grainy/spotty thing on the Ponceau might be too much SDS in the gel that gets transferred on the membrane... but I wouldn't put my money on that. If you indeed have more than normal amount of SDS in your gel, also the transfer would be worse than usual.
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Posted 05 September 2012 - 12:45 PM