Silver Destaining Using Hydrogen Peroxide
| Step | Repeats | Time | Solution | Purpose |
| 1 | 2X | 5 min | Water | Remove acid and PAGE buffers |
| 2 | 2X | 5 min | 25mM NH4HCO3 | pH adjustment and silver complexation |
|
3 |
1X | Until complete (<10 min) | 1% H2O2 in 25mM NH4HCO3 | Silver oxidation and removal |
| 4 | 2X | 5 min | Water | H2O2 and Ag+ removal |
| 5 | 1X | 5 min | 1% Formic acid | Lower pH of gel plug |
| 6 | 1X | 5 min | 50% Water: 50% acetonitrile w/1% Formic acid | Sequential dehydration of gel band/plug |
| 7 | 1X | 10 min | Acetonitrile | Dehydration of gel band/plug |
Recommended volumes are 25µL for 1mm diameter gel plugs and 50 µL for bands: however, these can be adjusted to meet need or personal preference
Reference:
[1] - Rapid Commun. Mass Spectrom. 2002; 16: 160-168
