SDS PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis)
|
Running
|
Stacking
|
7.5%
|
9%
|
10%
|
11%
|
12%
|
12.5%
|
15%
|
3%
|
10% SDS
|
0.3 ml
|
0.3 ml
|
0.3 ml
|
0.3 ml
|
0.3 ml
|
0.3 ml
|
0.3 ml
|
0.1 ml
|
40% Acrylamide
|
3.75 ml
|
4.5 ml
|
5 ml
|
5.5 ml
|
6 ml
|
6.25 ml
|
7.5 ml
|
0.75 ml
|
Running buffer
(3M Tris pH8.9)
|
2.5 ml
|
2.5 ml
|
2.5 ml
|
2.5 ml
|
2.5 ml
|
2.5 ml
|
2.5 ml
|
|
Stacking buffer
(0.5M Tris pH6.7)
|
|
|
|
|
|
|
|
1.25 ml
|
H2O
|
12.55 ml
|
10.77 ml
|
10.6 ml
|
8.77 ml
|
7.94 ml
|
7.52 ml
|
5.6 ml
|
7.8 ml
|
2.5M Sucrose
|
0.8 ml
|
1.83 ml
|
1.5 ml
|
2.83 ml
|
3.16 ml
|
3.33 ml
|
4 ml
|
|
TEMED
|
16.6 μl
|
16.6 μl
|
16.6 μl
|
16.6 μl
|
16.6 μl
|
16.6 μl
|
16.6 μl
|
16.6 μl
|
10% AP
|
83 μl
|
83 μl
|
83 μl
|
83 μl
|
83 μl
|
83 μl
|
83 μl
|
83 μl
|
Total volumn
|
20 ml
|
20 ml
|
20 ml
|
20 ml
|
20 ml
|
20 ml
|
20 ml
|
10 ml
|
* 最後要loading gel前才加AP，使gel凝固
Electrophoresis buffer（10 ×）：
60 g Tris，284 g Glycine，10 g SDS，final volume to 2 L with ddH2O.
將玻璃平整置於鑄膠台上(注意是否會漏)，接著將所需比例之running gel配置好後注入(1.5mm 約注入8ml)，上層注入H2O(維持膠體之平整並去除氣泡)，靜置等其凝固。
下層膠體凝固後將H2O移除，接著將配置好之stacking gel注入，並於上方插入所需樣品數之齒梳，靜置至上層凝固即可使用。
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