Chemiluminescence reagent

Tris-Cl (100 mM, pH 8.5)

hydrogen peroxide (30%)

Store at 4°C

Luminol (250 mM in DMSO)

Store at room temperature, wrapped in foil

p-Coumaric acid (90 mM in DMSO)

Store at room temperature, wrapped in foil

To 12 ml of Tris-Cl (100 mM, pH 8.5), add 3.8 μl of H₂O₂ (30%), 60 μl luminol (250 mM), and 26.6 μl p-coumaric acid (90 mM). This quantity is sufficient for a large blot. Prepare only as much reagent as needed because it has a very short shelf life.