Stacking Gel Polymerization
Posted 31 March 2011 - 09:22 PM
Posted 01 April 2011 - 05:46 AM
Posted 01 April 2011 - 07:09 AM
You could also try to use freshly made acrylamide, and verify the pH of your buffers.
Posted 04 April 2011 - 04:13 AM
My lower resolving gel for western blots has always turned out well. My upper, stacking gel, however, continues to fail me. When I insert the comb, there are no bubbles. When I come back to the gel 30 minutes later and it is polymerized, the gel has completely pulled away from the comb, ruining some of the wells. I understand that this is natural to an extent but sometimes only four of my wells are viable. Any suggestions to solve/manage the problem? How do I get the gel to 'shrink' less or at least in an upward fashion so the bottom of my wells are usable?
Try this protocol!!!!
2.9 ml H2O
1.25 ml Upper Tris
25ul APS (Please vortex APS when u take from -20 (u should mix well anything u take from -20/-70))
hope it will works..