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Coomassie Blue

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Silver stain

silver staining, silver stain

Silver development protocol (silver stain) for small 0.75mm polyacrylamide gels

1- Agitate gel twice with 50% MeOH, 10% acetic acid (Fixer 1, 100ml) for 15min ea.

2- Agitate in 10% EtOH, 5% glacial acetic acid (Fixer 2, 100ml) for 6min.

3- Rinse (1 min) with 100ml DW.

4- Agitate (7 min) with 100ml DW.

5- Agitate (11 min) with 100ml DW.

6- Agitate in 100ml from (250mlDDW with 5mg Na₂S₂O₄) (hydrosulfite [dithionite], make fresh) 9min.
********Add water to the hydrosulfite immediately before washing

7- Pour off solution; w/o rinsing add 100ml, 0.1% AgNO₃ (100mg) in DDW add 75ul 37% formaldehyde. Agitate 9min.

8- Rinse 30 sec with DDW to remove excess AgNO₃; add 100ml image developer (1ml 37% formaldehyde per liter of 3% sodium carbonate) mixed with 100ul 10g/L sodium thiosulfate, Agitate to desired staining intensity (3 to 6min).

9- Pour off developer, add 60 ml of stop solution (50g tris, 25ml glacial acetic per liter of distilled water).

10- Store gel in 10% glycerol with bacteriostat (0.05% NaN₃).