|
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
|
