Silver staining procedure

Chemicals

for 2 small. gels

for 4 small gels

for 6 small gels

Solution 1

Ethanol

Acetic Acid

40 ml

10 ml

80 ml

20 ml

120 ml

30 ml

Ad Milli-Q-H₂O up to

100 ml

200ml

300ml

Solution 2

Ethanol

45 ml

90ml

135 ml

Ad Milli-Q-H₂O up to

150 ml

300ml

450 ml

Solution 3

Na₂S₂O₃ x 5 H₂O

0,01g

0,02g

0,03 g

Ad Milli-Q-H₂O up to

50 ml

100ml

150 ml

Solution 4*
wrap with aluminium foil, lightsensitiv

AgNO₃

0,1 g

0,2g

0,3 g

Ad Milli-Q-H₂O up to

50 ml

100ml

150 ml

Solution 5 = Developing-Solution

Na₂CO₃

Na₂S₂O₃ x 5 H₂O = Solution 3

Formaldehyde (37%)

3 g

1,2 ml

67,5 µl

6g

2,5ml

135µl

9 g

3,7 ml

202,5 µl

Ad Milli-Q-H₂O up to

50 ml

100ml

150 ml

Solution 6
= STOP- Solution

EDTA (Stock 250 mM)

Acetic Acid

10 ml

0,5 ml

20ml

1 ml

30 ml

1,5 ml

Ad Milli-Q-H₂0 up to

50 ml

100ml

150 ml

Step

Time [min]

Solution

1

30

Solution 1: 40% EtOH, 10% Acetic Acid

2

30

Solution 1

3

20

Solution 2: 30% EtOH

4

20

Solution 2

5

20

Solution 2

6

1

Solution 3: 0,2 g/l Na₂S₂O₃ x 5H₂O

7

1

MilliQ H₂O

8

1

MilliQ H₂O

9

20

Solution 4: 2 g/l AgNO₃

10

1

MilliQ H₂O

11

1

MilliQ H₂O

12

Solution 5: 6% Na₂CO₃/5 mg/l Na₂S₂O₃ x 5H₂O/ 0,05% Formaldehyde

13

Solution 6: STOP solution