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SDS PAGE with BioRad Mini-Gel apparatus 1. Clean glass plates and spacers. Set up gel plates with BioRad clamps. Place in casting stand. Mark
 

Summary: 34
SDS PAGE with BioRad Mini-Gel apparatus
1. Clean glass plates and spacers. Set up gel plates with BioRad clamps. Place in casting stand. Mark
position of bottom of wells.
2. Prepare resolving gel. Combine all but APS and TEMED. Fresh APS = 0.25 g APS + 0.85 ml dH2O.
10.5% 2 X 10.5% 12%
5X 8.8 Buffer 0.8 ml 1.6 ml 0.8 ml
30% Acrylamide 1.4 ml 2.8 ml 1.6 ml
ddH2O 1.8 ml 3.6 ml 1.6 ml
TEMED 4 Ál 8 Ál 4 Ál
25% APS 8 Ál 16 Ál 8 Ál
Total Volume 4 ml 8 ml 4 ml
3. Seal plates: Combine 200 Ál resolving gel mix with 1 Ál TEMED + 1 Ál APS. Pour 100 Ál down
inside of each spacer. Allow to polymerize at bottom (1-2 minutes).
4. Add APS and TEMED to resolving gel mix (amounts can be increased 25%). Swirl gently. Pour to
~5 mm below bottom of wells (~3.5 ml). Overlay with dH2O-saturated butanol. Will polymerize
in 15-20 minutes
5. Pour off unpolymerized resolving gel mix. Wash with dH2O several times (squirt bottle). Place on
side to drain dH2O completely.
6. Prepare stacking gel. Use APS made above.

  

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

 

Collections: Biology and Medicine