When the reaction is stopped with tris base, the solution must be left at room temperature. At higher temperatures and at alkaline pH, colour "leaks" from the substrate. Also the enzyme may not be completely inactivated at pH 8.5 (as is the case for bacterial alpha-amylase). For bacterial alpha-amylase, we recommend the use of 2% tri-sodium phosphate (pH 11.0) as the stopping reagent.
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