Difference between revisions of "Preparing a SDS-PAGE Gel"
From Bridges Lab Protocols
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Revision as of 18:52, 3 August 2009
Separating Gel
- For 12 mL final volume (2 gels)
- Combine, adding APS and TEMED last
- Pour into cassette, overlay with the top layer of water saturated butanol
| Component | 7.5% | 10% | 12.5% |
|---|---|---|---|
| SDS-PAGE Separating Gel Buffer | 3 mL | 4 mL | 5 mL |
| Acrylamide Solution | 3 mL | 3 mL | 3 mL |
| Water | 6 mL | 5 mL | 4mL |
| APS (make fresh, 0.1g/mL in water) | 200 uL | 200 uL | 200 uL |
| TEMED | 30 uL | 30 uL | 30uL |
Stacking Gel
- Rinse polymerized separating gel with water
- For 10 mL (2 gels)
- Combine, adding APS and TEMED last
- Pour over separating gel and insert comb
| Component | |||
|---|---|---|---|
| SDS-PAGE Stacking Gel Buffer | 2.5 mL | ||
| Acrylamide Solution | 1.7 mL | ||
| Water | 4 mL | ||
| APS (make fresh, 0.1g/mL in water) | 200 uL | 200 uL | 200 uL |
| TEMED | 30 uL | 30 uL | 30uL |
