Editing Silver Staining

1. Fix<br>30% EtOH and 10% HOAc, 3 times 5 min each; 

2. Wash<br>30% EtOH, 3 times 5 min each; 

3. Rehydrate<br>MQ Water, 20 min; 

4. Decrease Background<br>Sensitizer, 2 min; 

5. Wash<br>MQ water, 2 times 5 min each; 

6. Expose to Silver<br>20ul Enhancer and 25ml Stainer, 30min; 

7. Wash<br>MQ water, about 1 min; 

8. Develop<br>watch closely to get dark enough<br>12.5ul Enhancer and 25ml Developer, Fast shaking with 200ul Sensitizer, 2-10 min 

9. Stop<br>try to anticipate how dark you want it and stop just before this point.<br>2.5 ml Acetic Acid; 

10. Wash with water. 

STOCK SOLUTIONS: 

Fixer for Silver: 150ml EtOH, 50ml HOAc, 300ml MQ water;<br>Washer: 15ml EtOH, 35ml MQ water;<br>Sensitizer: 2.5 mM Sodium Thiosulfate 0.105g/250ml (FRESH for 1week);<br>Stainer: 0.2% AgNO3 0.2 g/100 ml;<br>Developer: 3% Na2CO3 3g/100 ml;<br>Enhancer: 37% Formaldehyde; 

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== Sliver staining 2 ==

Fixer for Silver: 150 ml EtOH, 50 ml HOAc, 300 ml MQ water;<br>Protocol used by Liang in Bear Lab - UNC at Chapel Hill<br>Washer: 15 ml EtOH, 35 ml MQ water;<br>Sensitizer: 2.5 mM Sodium Thiosulfate 0.105 g/250 ml (FRESH for 1 week);<br>Stainer: 0.2% AgNO3 0.2 g/100 ml;<br>Developer: 3% Na2 CO3 3 g/100 ml;<br>Enhancer: 37% Formaldehyde;<br>1. Fix in 30% EtOH and 10% HOAc, 3 times 5 min each;<br>3. Rehydrate in MQ Water, 20 min;<br>2. Wash with 30% EtOH, 3 times 5 min each;<br>4. Decrease background with Sensitizer, 2 min;<br>5. Wash with MQ water, 2 times 5 min each;<br>6. Expose to Silver, using 20 μl Enhancer and 25 ml Stainer, 30 min;<br>7. Wash with MQ water, about 1 min;<br>8. Develop for 2-10 min (watch closely to get dark enough) using 12.5 μl Enhancer and 25 ml Developer mixed with 200 μl Sensitizer;<br>9. Stop by 2.5 ml acetic acid (try to anticipate how dark you want it and stop just before this point);<br>10. Wash with water.<br><br>

[[Category:Protocols]]