step

reagent 

volume

gel>0.5 mm

1 Fixing

20% trichloroacetic acid (w/v)
(40 ml “100%” TCA-solution in 200 ml)

200 ml

45 min

2 Washing

H₂O dist.

200 ml

5 min

3 Rinsing #1

50% ethanol/ 10% acetic acid (v/v)
(100 ml EtOH / 20 ml HAc in 200 ml)

200 ml

40 min

4 Rinsing #2

5% ethanol/ 7% acetic acid (v/v)
(10 ml EtOH / 14 ml HAc in 200 ml)

200 ml

20 min

5 Incubation

165 mM Sodium Acetate; 2.5 % Glutardialdehyde (w/v)
0.07% (w/v) Benzenesulfonic acid
(4.5 g NaAc / 20 ml Benzenesulfonic stock sol.,
20 ml Glutardialdehyde (25%) in 200ml) 

200 ml

20 min

6 Washing

H₂O dist.

3 x 200 ml

3 x 20 min

8 Silvering

0.2% AgNO₃ /0.05% Formaldehyde (w/v), 0.07% Benzenesulfonic Acid
(8 ml AgNO_{3 sol.,} 20 ml Benzenesulfonic acid sol., 260ul HCHO (37% w/v) per 200 ml)

200 ml

40 min

9 Washing

H₂O dist.

3 x 200 ml

3 x 1 min

10 Developing

2.5 % Na₂CO₃ / 0.05% Formaldehyde
(5g Na₂CO₃, 260ul HCHO (37% w/v) per 200 ml)

200 ml

2 -3 min

11 Stopping
Preserving

10% HAc, 5% Glycerol
(20 ml HAc + 10 ml Glycerol)

200 ml

30 min

Drying

air dry , on the support-film

---

overnight