Glutamate Oxidase Assay Kit (Fluorometric - Near Infrared)
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Functional Studies - Glutamate Oxidase Assay Kit (Fluorometric - Near Infrared) (AB138885)
Sample Standard Curve for Glutamate Oxidase. Glutamate oxidase dose response was measured on a 96- well black plate with ab138885. As low as 40 µU/mL glutamate oxidase can be detected with 60 minutes incubation time (n=3).
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Product details
Glutamate Oxidase Assay Kit (Fluorometric - Near Infrared) (ab138885) provides a quick and an ultrasensitive method for the measurement of glutamate oxidase in solution and in cell lysates. In the assay, L-glutamic acid is oxidized to α-ketoglutarate, NH3 and H2O2 by glutamate oxidase. The kit uses our AbRed Indicator substrate which can react with H2O2 when catalyzed by horseradish peroxidase (HRP) to generate the highly fluorescent product. The signal can be read with either a fluorescence microplate reader at Ex/Em = 530-570 nm/590 -600 nm (optimal Ex/Em = 540 nm/590 nm) or an absorbance microplate reader at 576±5 nm. With this product, 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.
Glutamate oxidase specifically catalyzes the oxidative deamination of L-glutamate in the presence of water and oxygen with the formation of ketoglutarate, ammonia, and hydrogen peroxide. Glutamate oxidase can be used as an analytic reagent and as a basis for developing biosensors for the determination of L-glutamate, L-glutamine, ammonia, and creatinine. These biosensors can be used in clinical biochemistry for the determination of glutamate-pyruvate transaminase and glutamate-oxalacetate transaminase in biological fluids, which makes the early diagnosis of heart and liver diseases possible.
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Biological function summary
Glutamate oxidase plays a significant role in nitrogen metabolism. It functions effectively by breaking down the amino acid glutamate playing a role in cellular energy processes. By generating hydrogen peroxide as a by-product the enzyme contributes to redox balancing within the cell. It acts independently not forming extensive complexes with other proteins which maintains its specificity to its substrates and reactions.
Pathways
Glutamate oxidase integrates with cellular respiratory and metabolic pathways. It links to the tricarboxylic acid (TCA) cycle through its production of α-ketoglutarate an important TCA cycle intermediate. The enzyme shares functional intersections with proteins like glutamine synthetase which recycles ammonia back into glutamine highlighting its place in amino acid and nitrogen metabolism.
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Publications (1)
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Talanta 141:175-81 PubMed25966399
2015
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