SDS - Polyacrylamide Gels
Separation gel:
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| Gel % | 6.0 | 7.5 | 8.0 | 10 | 12 | 6.0 | 7.5 | 8.0 | 10 | 12 | |
| 4 x lower buffer (in ml) | 2.5 | 2.5 | 2.5 | 2.5 | 2.5 | 5.0 | 5.0 | 5.0 | 5.0 | 5.0 | |
| 40 % (30 : 0.8) acrylamide/ bisacrylamide (in ml) |
1.5 | 1.9 | 2.0 | 2.5 | 3.0 | 3.0 | 3.8 | 4.0 | 5.0 | 6.0 | |
| H2O (in ml) | 6.0 | 5.6 | 5.5 | 5.0 | 4.5 | 12.0 | 11.2 | 11.0 | 10.0 | 9.0 | |
| APS (25 %) in µl | 30 | 30 | 30 | 30 | 30 | 60 | 60 | 60 | 60 | 60 | |
| TEMED in µl | 15 | 15 | 15 | 15 | 15 | 30 | 30 | 30 | 30 | 30 | |
5 % stacking gel:
| for 5 ml | for 10 ml |
| 4 x upper buffer (in ml) | 1.25 | 2.5 |
| 40 % (30 : 0.8) acrylamide/ bisacrylamide (in µl) |
625 | 1250 |
| H2O (in ml) | 3.1 | 6.25 |
| APS (25 %) in µl | 30 | 60 |
| TEMED in µl | 15 | 30 |
Buffers:
4 x lower buffer: 1.5 M Tris/HCl pH
8.8; 0.4% SDS.
[for 500ml: 91 g Tris/HCl; 2 g SDS]
4 x upper buffer: 0.5 M Tris/HCl pH
6.8; 0.4% SDS.
[for 500ml: 30.4 g Tris/HCl; 2 g SDS]
10 x running buffer: 0.25 M Tris;
2 M Glycin; 1 % SDS.
[for 1 l: 30.4 g Tris; ; 150 g Glycin; 10 g SDS]
2 x loading buffer (reducing): 80
mM Tris/HCl pH 6.8; 10 % Mercaptoethanol;
2 % SDS; 10 % Glycerin; Bromphenol Blue.
[for 10 ml: 800 µl 1 M Tris/HCl pH 6.8; 1 ml Mercaptoethanol;
2 ml 10 % SDS; 2 ml 50 % Glycerin, Bromphenol Blue]
