small proteins have run ahead of tracking dye
Posted 12 May 2010 - 09:24 AM
I run an acrylamide gel electrophoresis for RIPA and have encountered a new problem after preparing some new reagents. The smaller mol. weight proteins have run off the gel as if I let it run long, but the tracking dye had only just reached the bottom of the gel. Tracking dye was one of the new reagents. Any other likely candidates? I run at constant power (amps) and the voltage started and finished a little lower than usual - usually starts at ~107v and runs up to ~390V after 4 hours, but this time 97v to 335v after 4.5 hrs. I also decided to make my stacking gel a little taller - usually 1cm from well bottom to top of resolving gel, this time 2cm, but I've run it this way before without the run-off problem.
Thanks for any advice...
Posted 18 May 2010 - 11:08 AM
check the pH of all of your buffers.
lower voltage indicates that the ionic strength is higher. the buffers may be made wrong (too strong).
these factors may affect mobility.
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genius does what it must
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