The Larochelle Lab
at Clark University

...................................SILVER STAINING


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


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


Dry at 70-80°C for 1-2 hours.

For a 1.5 mm gel, all incubations should be twice as long.


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Clark Biology Dept | Clark University

Worcester, MA | 508.793.7631 |