Amplite(TM) Fluorimetric Glutamate Oxidase Assay Kit *Red Fluorescence*

Glutamate oxidase belongs to the family of oxidoreductases, specifically those acting on the CH-NH2 group of donors with oxygen as an acceptor. It is an enzyme that specifically catalyzes the oxidative deamination of L-glutamate in the presence of water and oxygen with the formation of o-ketoglutarate, ammonia, and hydrogen peroxide. The Amplite(TM) Fluorimetric Glutamate Oxidase Assay Kit provides a quick and ultrasensitive method for the measurement of glutamate oxidase in solution and in cell lysates. In the assay, L-glutamic acid is oxidized to o-ketoglutarate, NH3 and H2O2 by glutamate oxidase. L-Alanine and L-glutamate-pyruvate transaminase are included in the reaction, resulting in multiple cycles of the initial reaction, thus significantly amplifying the production of H2O2. The kit uses our Amplite(TM) Red substrate which enables a dual recordable mode. The fluorescent signal can be easily read by either a fluorescence microplate reader at Ex/Em = 540/590 nm or an absorbance microplate reader at 576 nm. With the Amplite(TM) Glutamate Oxidase Assay kit, we have detected as little as 40 µU/mL glutamate oxidase in a 100 µL reaction volume. It can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.