Brief Description
Gel percentages and instructions for pouring gels
Protocol Details
Western Blot Gel Set-up
SDS-PAGE GELS
Separating Gel
|
% |
5 |
6 |
7 |
7.5 |
8 |
9 |
10 |
12 |
13 |
15 |
16.5 |
|
30% acrylamide |
1.67 |
2 |
2.33 |
2.5 |
2.67 |
3 |
3.33 |
4 |
4.33 |
5 |
5.5 |
|
separating buffer |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
2.5 |
|
ddi H2O |
5.83 |
5.5 |
5.17 |
5 |
4.83 |
4.5 |
4.17 |
3.5 |
3.17 |
2.5 |
2 |
10% APS 50uL
TEMED 12uL
Stacking Gel
| 30% acrylamide | .67mL |
| stacking buffer | 1.25mL |
| ddi H20 | 3mL |
10% APS 30uL
TEMED 7uL
Note: above volumes are for two gels
-Add 10% APS and TEMED just before pouring gel
-Pour separating gel, ~5mL /gel and add a small volume of t-butanol or 2-propanol and let stand ~15 minutes until gel polymerizes
-After gel polymerizes, pour off t-butano. Remove excess with P200 pepette and paper towel
-Pour stacking gel and let stand until polymerized