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Silver Stain Protocol

Oksana’s Fantastic Silver Staining Method


Solutions: (make everything up fresh):

Fixer: 60 ml 50% Acetone

1.5 ml 50% TCA

25 µl Formaldehyde (37%)


Pre-Stain: 10 mg Na thiosulphate

in 60ml dH2O


Stain: 160 mg Silver nitrate

600 µl Formaldehyde

in 60 ml dH2O


Developer: 1.2 g Na2CO3

25 µl Formaldehyde

2.5 mg Na thiosulfate

in 60 ml dH2O


Stopping Solution: 1% Acetic Acid (can store)


Others: 50% Acetone (can store)


Method:

1. Put gel in fixer for 15 mins (shaking)

2. Wash with dH2O: 1 3x quick wash,

1 1x 5 mins,

1 3x quick

3. Wash with 50% acetone for 5 mins

4. Pour off and add pre-stain for 1 min

5. Wash with dH2O 1 3x quick

6. Stain for 8 mins

7. Wash with dH2O 1 2x quick

8. Develop for 10-20 sec (Take care as bands appear very quickly!)

9. Remove developer and add stopping solution for 1-2 mins

10. Rinse with dH2O and leave in water until ready to dry