|The Glutamate n/a (Catalog #MBS169257) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase.
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Please refer to the product datasheet for known applications of a given assay kit. We\'ve tested the Glutamate Assay Kit with the following immunoassay(s):
Typical Testing Data/Standard Curve
Principle of the Assay: The Glutamate Assay Kit is a sensitive quantitative fluorometric assay for glutamate. Glutamate oxidase converts glutamate to alpha-ketoglutarate and also produces NH3 as well as H2O2. L-alanine and glutamate-pyruvate transaminase are also added to the reaction in order to regenerate glutamate. As a result, multiple rounds of the reaction occur which results in significant amplification of H2O2 production. In the presence of HRP, the Fluorometric Probe reacts with H2O2 in a 1:1 stoichiometry to produce highly fluorescent Resorufin. The Resorufin product can be easily read by a fluorescence microplate reader with an excitation of 530-560 nm and an emission of 590 nm. Fluorescence values are proportional to the glutamate levels within the samples. The glutamate content in unknown samples is determined by comparison with a standard curve.
Background/Introduction: Glutamate is a non-essential amino acid that has a key metabolic role in processes such as the citric acid cycle and removal of excess nitrogen waste. In its monosodium form (MSG), glutamate is well known as a flavor enhancer. Glutamate has also been identified as one of the major excitatory neurotransmitters of the mammalian brain. Glutamate is involved in learning and memory, and long term potentiation occurs at glutaminergic synapses. In addition, glutamate helps to regulate growth cones and synaptogenesis. Postsynaptically, glutamate has been suggested to activate the NMDA, AMPA, and kainite receptors. Damage and/or death to nerve cells due to excessive glutamate release and deficits in uptake have been correlated with diseases such as amyotrophic lateral sclerosis, lathyrism, and Alzheimer\'s disease as well as stroke, autism, and some forms of intellectual disability. Glutamate Assay Kit is a simple HTS-compatible assay for measuring glutamate levels in biological samples without any need for pretreatment. The assay uses glutamate-specific enzymes to generate H2O2. In the presence of H2O2 and horseradish peroxidase (HRP), the non-fluorescent Fluorometric Probe is oxidized to the highly fluorescent Resorufin. The kit has a detection sensitivity limit of 300 nM glutamate. Each kit provides sufficient reagents to perform up to 200 assays, including standard curve and unknown samples.