Cereal alpha-amylase assay reagent cannot be used at pH 3-4. However, you can perform a similar assay using blocked p-nitophenol maltoheptaoside (BPNPG7). Basically dissolve the BPNPG7 vial contents in 10 mL with water. Incubate 0.2 mL of your enzyme (buffered) with 0.2 mL of BPNPG7 for 10 min at a set temperature. Terminate the reaction by heating in a boiling water bath for 2 min. Then add 0.2 mL of alpha-glucosidase (E-TSAGL; at 10 U/mL in 0.2 M tris buffer, pH 7.0) and incubate for 10 min. Add 3 mL stopping reagent (Trizma base pH 8.5) and read colour at 400 nm.
Any recommendations on using your α-Amylase Assay Kit (Ceralpha Method) (K-CERA) assay at a low pH (pH 3-4)?
Modified on: Tue, 29 Mar, 2016 at 2:22 PM
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