
Stacking Gel Polymerization
#1
Posted 31 March 2011 - 09:22 PM
#2
Posted 01 April 2011 - 05:46 AM
#3
Posted 01 April 2011 - 07:09 AM
You could also try to use freshly made acrylamide, and verify the pH of your buffers.
#4
Posted 01 April 2011 - 07:14 AM
#5
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?
Hi,
Try this protocol!!!!
for 5ml:
2.9 ml H2O
1.25 ml Upper Tris
0.85ml Acrylamide
25ul APS (Please vortex APS when u take from -20 (u should mix well anything u take from -20/-70))
10ul TEMED
hope it will works..
gud luk...