| Percentage of Gel | 5.0 | 7.5 | 10.0 | 12.5 | 15.0 | 17.5 | 20.0 |
| 30% w/v Acrylamide | 83 | 125 | 167 | 208 | 250 | 292 | 333 |
| 1% w/v bis-acrylamide | 130 | 97 | 65 | 52 | 43 | 37 | 33 |
| 1.5 M Tris-HCl pH 8.8 | 125 | 125 | 125 | 125 | 125 | 125 | 125 |
| Water | 159 | 150 | 140 | 112 | 79 | 43 | 6 |
| Final Ratio Acryl:Bis | 19:1 | 38.6:1 | 77:1 | 120:1 | 174:1 | 237:1 | 303:1 |
Add 50 ul of 10% ammonium persulfate, degas under vacuum, add 5 ul of TEMED per 10 ml of mix.
2. Stacking Gel Mix
| 30% w/v acrylamide | 25.0 | (5% final) |
| 1% w/v bisacrylamide | 20.0 | (0.13% final) |
| 1 M Tris-Cl pH 6.8 | 18.75 | (125 mM final) |
| Water | 86.25 | (to make 150 ml total) |
Add 50 ul of 10% ammonium persulfate and 5 ul of TEMED per 5 ml of mix.
3. Sample Buffer (store at Room Temp)
| 10% w/v SDS | 30.0 | (2% final) |
| 1 M Tris-Cl pH 6.8 | 12.0 | (80 mM final) |
| 50% v/v Glycerol | 30.0 | (10% v/v final) |
| Bromophenol Blue | to taste | (try 1 ml of 0.2% stok in EtOH) |
| Water | 78.0 | (to make 150 ml total) |
4. Running Buffer
| Glycine | 288 g |
| Tris Base | 60 g |
| 10% w/v SDS | 100 ml |
| Water | to 10 litres |
5. Stain
| 5 g | Coomassie R-250 |
| 450 ml | Methanol |
| 100 ml | Glacial Acetic Acid |
| 450 ml | Water |
dissolve R-250 in methanol first, then add other solvents.
5. De-stain
25% v/v Methanol, 7% Acetic Acid