The α-Amylase HR Assay Reagent (R-AMHR4) cannot be used at pH 3-4. However, you can perform a similar assay using blocked p-nitrophenyl maltoheptaoside (O-BPNPC7). Basically dissolve the BPNPC7 vial contents in 10 mL of water. Incubate 0.2 mL of your enzyme (buffered) with 0.2 mL of BPNPC7 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.
Modified on: Tue, 1 Oct, 2019 at 10:15 AM
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