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Western Blotting: Running at too high of a voltage?

Western Blotting: Running at too high of a voltage? - Western Blot Forum

Western Blotting: Running at too high of a voltage? - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 04-09-2010, 07:19 PM
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Default Western Blotting: Running at too high of a voltage?



Hi, I've been getting some streaking in my marker lanes and to a limited extent with my sample lanes. The protocol my PI wants me to use has me running at 200V with minigels at benchtop temperature. Is it possible that this is what's giving me streaking? Should I lower my voltage or maybe just move my setup into the cold room?
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Old 04-12-2010, 06:02 PM
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Default Re: Western Blotting: Running at too high of a voltage?

You can minimize streaking by lowering the voltage of the gel run. No big deal. You should run it longer until the dye front reaches the end of the gel.
It will probably help, because it will give the electrophoresis time to remove excess salts (if any) in your sample.
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