CHP
Fix 3 * 5 min 2* 45 min or overnight
30% EtOH / 10% HOAc 300ml/L & 100ml/L
1 __________ 2__________
Wash 3* 15 min or 3X 5 min
30% EtOH 1 __________ 2 _________ 3 _________ 300ml/l
Rehydrate 1* 20 min dd Water __________
Decrease Background
1* 2 min (for 1.5 mm gels: 4 min)
2.5 mM NaThiosulfate (0.62 g/L) 0.31 g/500ml
Time: ____________
Wash 2* 5 min dd Water 1 _________ 2 __________
Expose to Silver
1* 30 min 0.2% AgNO3 0.2 g/100 ml
0.8 ml Formaldehyde 37% per liter 80 ul /100ml
Time:__________
*** Dispose of in the silver waste container ***
Wash 1* 1 min dd Water
Revelation
2-10 min 3 % Na2CO3 - Lot# 3g/100 ml
0.5 ml formaldehyde 37% 50ul /100ml
20uM NaThiosulfate (8ml of 2.5 mM) 800ul/100ml
per liter Time:
Stop 10 ml Acetic Acid
Try to anticipate how dark you want it and stop just before this point.
Wash dd Water