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.
Was this article helpful?
That’s Great!
Thank you for your feedback
Sorry! We couldn't be helpful
Thank you for your feedback
Feedback sent
We appreciate your effort and will try to fix the article