Small SDS-PAGs (7.5 x 10 cm gels of 0.75-1 mm thickness)
Resolving gel (10 ml for 2 gels):
|
|
5% |
10% |
12% |
15% |
20% |
Lower buffer |
2.5 ml |
2.5 ml |
2.5 ml |
2.5 ml |
2.5 ml |
Acrylamide, 30% |
1.6 ml |
3.3 ml |
4 ml |
5 ml |
6.6 ml |
|
H2O |
5.9 ml |
4.2 ml |
3.5 ml |
2.5 ml |
0.9 ml |
TEMED |
10 μl |
10 μl |
10 μl |
10 μl |
10 μl |
|
10% APS |
100 μl |
100 μl |
100 μl |
100 μl |
100 μl |
Stacking 3.3% gel (5 ml for 2 gels):
Upper buffer 1.25 ml Resolving gel: 0.375 M Tris final
Acrylamide, 30% 0.55 ml Stacking gel: 0.15 M Tris final
H2O 3.2 ml 10% APS: 0.1% final
TEMED 5 μl SDS: 0.1 and 0.125% final
10% APS 50 μl
Lower buffer, pH 8.8:
-
1.5 M Tris (’Sigma’, T-1378, Fw 121.1 g/mol)/HCl => 18.165 g/100 ml H2O
-
0.4% SDS (’BDH’, No. 44244, Fw 288.38 g/mol) => 0.4 g
Upper buffer, pH 6.8:
-
0.6 M Tris/HCl => 7.266 g/100 ml H2O
-
0.5% SDS => 0.5 g
2003 10 21 – Oliver & Julius