Examples of polyacrylamide gel preparation for SDS-PAGE are summarized by each gel concentration. Please use an acrylamide/bis mixed solution with an acrylamide/bis ratio of 37.5:1 or 29:1.
| For 10 mL of separation gel | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Gel concentration (%T) | 6% | 8% | 10% | 12% | 15% | ||||||
| Concentration of acrylamide/bis mixed solution | 30w/v% | 40w/v% | 30w/v% | 40w/v% | 30w/v% | 40w/v% | 30w/v% | 40w/v% | 30w/v% | 40w/v% | |
| Product Number | Product Name | ||||||||||
| 043-16785 | Water(mL) | 5.4 | 5.9 | 4.7 | 5.4 | 4.1 | 4.9 | 3.4 | 4.4 | 2.4 | 3.65 |
| - | Acrylamide/bis mixed solution (mL) | 2.0 | 1.5 | 2.7 | 2.0 | 3.3 | 2.5 | 4.0 | 3.0 | 5.0 | 3.75 |
| 192-11041 | Separating Gel Buffer Solution (x 4) | 2.5 | 2.5 | 2.5 | 2.5 | 2.5 | 2.5 | 2.5 | 2.5 | 2.5 | 2.50 |
| 019-15922 | 10w/v% APS(mL) | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.10 |
| Total volume (mL) | 10.0 | 10.0 | 10.0 | 10.0 | 10.0 | 10.0 | 10.0 | 10.0 | 10.0 | 10.00 | |
| 208-17452 | TEMED(mL) | 0.008 | 0.008 | 0.006 | 0.006 | 0.004 | 0.004 | 0.004 | 0.004 | 0.004 | 0.004 |
* Deaeration is recommended before addition of TEMED.
* When TEMED is added, the gel starts to polymerize. Perform subsequent operations quickly.
| 5% concentrated gel | |||||
|---|---|---|---|---|---|
| Concentration of acrylamide/bis mixed solution | 30w/v% | 40w/v% | 30w/v% | 40w/v% | |
| Product Number | Product Name | For 1 mL | For 5 mL | ||
| 043-16785 | Water(mL) | 0.690 | 0.735 | 3.490 | 3.695 |
| - | Acrylamide/bis mixed solution(mL) | 0.170 | 0.125 | 0.830 | 0.625 |
| 199-11051 | Concentrated Gel Buffer Solution (x 4) | 0.130 | 0.130 | 0.630 | 0.630 |
| 019-15922 | 10w/v% APS(mL) | 0.010 | 0.010 | 0.050 | 0.050 |
| Total(mL) | 1.000 | 1.000 | 5.000 | 5.000 | |
| 208-17452 | TEMED(mL) | 0.001 | 0.001 | 0.005 | 0.005 |
Modified from Joseph and David. Cold Spring Harb Protoc; 2006.