SDS PAGE (Laemmli)
Materials
1. Acrylamide (29+1) %
2. x 4 lower gel buffer, 1.5M Tris-HCl pH 8.5, 0.4 % SDS
3. x 4 upper gel buffer, 0.5 M Tris-HCl pH 6.5, 0.4 % SDS
4. 10 % APS
5. TEMED
Lower gel
| x4 | A.A. | DW | ASP | TEMED | |
| 20% | 2.5 | 6.7 | 0.8 | 0.05 | 0.05 |
| 15% | 2.5 | 5.0 | 2.5 | 0.05 | 0.05 |
| 12.5% | 2.5 | 4.2 | 3.3 | 0.05 | 0.05 |
| 10% | 2.5 |
3.3 | 4.2 | 0.05 | 0.05 |
| 7.5% | 2.5 | 2.5 | 5.0 | 0.05 | 0.05 |
| 6% | 2.5 | 2.0 | 5.5 | 0.05 | 0.05 |
Upper gel
1. x 4 upper buf, 1.0
2. 30% A.A. 0.55
3. DW 2.35
4. APS 0.05
5. TEMED 0.05
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Total 4.0 ml
1 x Running buffer
Tris: 6.1 g
Glysin 28.8 g
10% SDS 20 ml
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Total 2000 ml
Sample buffer
10% glecerol 50%: 20.0
5% 2ME 2.5
2.5% SDS 10%: 12.5
0.125 M Tris-HCl pH6.7 1M: 6.25
DW: 8.75
Transfer buffer
Tris: 4.8 g
Glysin: 18.0 g
EtOH 400 ml
10% SDS 20 ml
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Total 2 l