...................................SILVER STAINING
Fixer: Developer: PROTOCOL: Links | About Prof Larochelle
20 mL dH2O
25 mL methanol
5 mL acetic acid
1.5 g Na2CO3
125 uL formaldehyde
250 mL dH2O
Put (0.75 mm) gel into fixer and rotate 15 minutes.
Wash 3x with dH2O, 5 minutes each wash.
Add 50 mL dH2O with 3.5 uL of 1M DTT and rotate 10 minutes.
Add 50 mg AgNO3 and 50 mL dH2O and rotate 10 minutes.
Add developer in about 5 portions. The first portion should turn
cloudy/grey suddenly. When this occurs, pour it off and replace with more
developer. Wait until either the developer appears to be cloudy or no
changes in staining occur before adding the remaining 3 portions of
developer.
Stop in 5% acetic acid for 5-10 minutes.
Put gel into 3% glycerol. Wet gel drying paper in 3% glycerol and lay flat
on gel dryer.
Place gel on paper and fold over the rest of it, trapping as few bubbles as
possible.
Dry at 70-80°C for 1-2 hours.
For a 1.5 mm gel, all incubations should be twice as long.
Clark Biology Dept | Clark University
Worcester, MA | 508.793.7631 | dlarochelle@clarku.edu