New Fuchsin Alkaline Phosphatase Substrate Solution – Red

Final Dilution:

        0.01% New Fuchsin, 0.02% Sodium Nitrite, 0.05% Naphthol AS-BI Phosphate, 1mM Levamisole, in 0.05M Tris-HCl, pH 8.7

Stock Solutions:

        0.2% New Fuchsin (20x) in 2N HCl and store at 4 °C.

        0.4% Sodium Nitrite (20x) and 20mM Levamisole (20x) in distilled water. Store at 4 °C.

        1% Naphthol AS-BI Phosphate (20x) (Sigma N2250) in 100% DMF (dimethylformamide) and store at 4 °C.

Working Solution:

       To 5 ml of 0.05M Tris-HCl, pH8.7,

       Add 5 drops of 0.2% New Fuchsin and mix well.

       Add 5 drops of 0.4% Sodium Nitrite/20mM Levamisole and mix well.

       Add 5 drops 1% Naphthol AS-BI Phosphate and mix well.

       Incubate sections for 10-20 minutes at room temperature. Rinse in tap water, counterstain and coverslip with permanent mounting medium.