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.
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- Talanta 141:175-812015Glutamate detection by amino functionalized tetrahedral amorphous carbon surfaces.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEmilia Kaivosoja et. al.PubMed 25966399
Key facts
- Detection method
- Colorimetric/Fluorometric
- Sample types
- Cell Lysate
- Assay type
- Enzyme activity
- Reactive species
- Mammals
- Sensitivity
- = 40 µU/mL
What's included?
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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.
Key facts
- Detection method
- Colorimetric/Fluorometric
- Sample types
- Cell Lysate
- Assay type
- Enzyme activity
- Reactive species
- Mammals
- Assay Platform
- Microplate reader
- Sensitivity
- = 40 µU/mL
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Glutamate oxidase sometimes called GluOx catalyzes the oxidative deamination of glutamate to form hydrogen peroxide ammonia and α-ketoglutarate. With a mass of approximately 62 kDa this enzyme facilitates the conversion of glutamate through a reaction that requires oxygen. It is predominantly expressed in microbial organisms such as fungi especially in Aspergillus species. In industrial settings scientists harness its ability to detect glutamate levels owing to its specificity and sensitivity in enzymatic reactions.
- 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.
- Associated diseases and disorders
Glutamate oxidase's activity becomes significant in neurological disorders. For example elevated glutamate levels associate with neurodegenerative diseases like Alzheimer's due to excitotoxicity. Additionally its role in producing hydrogen peroxide connects it to oxidative stress-related conditions such as amyotrophic lateral sclerosis (ALS). The enzyme intersects with proteins like superoxide dismutase 1 which mitigates oxidative damage highlighting the enzyme’s involvement in redox homeostasis.
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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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