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SDS PAGE with BRL Gel apparatus 1. Clean glass plates and spacers. Set up gel plates with clamps.
 

Summary: 35
SDS PAGE with BRL Gel apparatus
1. Clean glass plates and spacers. Set up gel plates with clamps.
2. Prepare resolving gel. Combine all but APS and TEMED. Degas for 2-4 minutes with H20 aspirator
(turn on water before attaching flask; turn off water after detaching flask). Use clean stopper.
10.5% 2 X 10.3%
5X 8.8 Buffer 4 ml 7 ml
30% Acrylamide 7 ml 12 ml
ddH2O 9 ml 16 ml
TEMED 20 Ál 35 Ál
25% APS 20 Ál 35 Ál
Make fresh APS (0.25 g APS + 0.85 ml ddH2O).
3. Seal plates. Combine: 1 ml SDS-PAGE mix + 5 Ál TEMED + 5 Ál APS. Spread along insides of
spacers. Place plates in horizontal position. Allow to polymerize at bottom (1-2 minutes).
4. Add APS and TEMED. Swirl gently. Pour ~15 ml gel. Overlay with butanol (ddH2O saturated).
5. Pour liquid off polymerized gel. Wash with ddH2O 3-5 times.
6. Prepare stacking gel. Use APS made above. Degas as above.
4.2%
5X 6.8 Buffer 1 ml
50% Sucrose 1 ml

  

Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida

 

Collections: Biology and Medicine