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Silver destaining using Hydrogen peroxide and ammonium bicarbonate [1]
|
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 |
[1] - Rapid Commun. Mass Spectrom. 2002; 16: 160-168
