AMPD Assay Kit is the first non-radioactive and continuous kit designed to measure AMP deaminase content in samples. This enzymatic assay is based on a reaction involving Inosine Monophosphate Dehydrogenase (IMPDH).
The principle of the assay is based on the coupling of the following enzymatic reactions:
(1) In the presence of AMP, AMP Deaminase enzyme catalyzes the formation of IMP.
(2) In the presence of NAD, IMP is immediately oxidized by a highly active IMPDH in the presence of NAD with simultaneous formation of NADH2 directly monitored spectrophotometrically at 340 nm.
The assay is developed for measuring AMPDA activity in vitro or in cell lysates.
For maximal accuracy, the assays with cell lysates are run with and without AMP in parallel. The absorbance rate observed in the absence of AMP is used as blank and is subtracted from the absorbance rate measured in its presence.