EASY AS 1-2-3 PROTOCOL
1. FIX gel for at least 25 minutes in 100ml of the standard mixture of 5:5:1 Methanol:Water:Acetic Acid. Decant fixative, then add reconstituted STERLING Fixative (45ml water, 50ml methanol, 5ml STERLING Fixative Concentrate) and fix for an additional 5 minutes
2. WASH gel twice for 15 minutes in deionized water. Addition of 0.1% non-ionic surfactant will aid in submerging the gel.

• While gel is washing, prepare Staining Solution as directed. Do not combine the two component solutions until just prior to use.

3. STAIN: Decant wash solution and immerse gel in combined staining solution Bands will begin to appear in 5-10 minutes. When desired intensity is achieved, stop development by immersing the gel in a 5% acetic acid solution.

Preparation of Staining Solution:
1. Dilute 25ml Reagent A with 25ml of water.
2. Dissolve 2.8 grams of Reagent B in 50ml of water. Stir until completely dissolved (approx. 5-10 minutes).
3. Immediately before use, pour (1) into (2) with stirring, and pour over gel. The combined solution has a useful life of ~20 minutes.