Glutamate Colorimetric Assay Kit

Catalog #: K629 | abID: ab83389

Product Details

abID ab83389
Cat # +Size K629-100
Size 100 assays
Detection Method Absorbance (450 nm)
Species Reactivity All
Applications N/A
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• Kit contains the necessary reagents for accurate measurement of Glutamate
Kit Components • Glutamate Assay Buffer
• Glutamate Enzyme Mix
• Glutamate Developer
• Glutamate Standard (0.1M)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Glutamate, one of the two acidic proteinogenic amino acids, is also a key molecule in cellular metabolism. In humans, glutamate plays an important role both in amino acid degradation and disposal of excess or waste nitrogen. Glutamate is the most abundant swift excitatory neurotransmitter in the mammalian nervous system. It is believed to be involved in learning and memory and has appeared to be involved in diseases like amyotrophic lateral sclerosis, lathyrism, autism, some forms of mental retardation and Alzheimer's disease. Glutamic acid is also present in a wide variety of foods, and has been used as a flavor enhancer in food industry. BioVision’s Glutamate Assay Kit provides a sensitive detection method of the glutamate in a variety of samples. The glutamate Enzyme Mix recognizes glutamate as a specific substrate leading to proportional color development. The amount of glutamate can therefore be easily quantified by colorimetric (spectrophotometry at λ = 450 nm) method.

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You mentioned the standard used in the kit is monosodium glutamate and the molecular weight of monosodium glutamate is 169.11 g/mol. However, the molecular weight given in the datasheet is for glutamate which is 147.13 g/mol. Why is the molecular weight of monosodium glutamate not used for calculations?
In biological samples, it is more appropriate to use the molecular weight of the anion (Glutamate - MW: 147.13 g/mol). If you’re calculating molarity from the Standard Curve, the molecular weight of the Standard doesn’t matter, as only the molecules of glutamate is counted. If you’re determining mg glutamate per volume, you have to use the glutamate molecular weight and not the molecular weight of monosodium glutamate.

To understand this better, any positive ion can be the counter-ion to the glutamate anion. Even a protein with molecular weight 1,000,000 g/mol with a positive charge can perform the function. However, it is meaningless to say that the molecular weight is 1,000147.13 g/mol for the combined molecule as the only part we need is the glutamate itself.
Can this assay only measure free glutamate or can it also measure the amount of glutamic acid in peptides/proteins?
This kit measures free glutamate levels only and not glutamic acid in the backbone of peptides/proteins.
Does glutamine cross-reat during this assay?
This assay uses enzymatic detection which use glutamate and not glutamine.
While using the Glutamate Assay kit to detect glutamate level in Astrocyte cell culture there is an increasing level of glutamate in culture, without adding any glutamate or without changing the culture medium. Why is this happening?
Astrocytes release glutamate in a Ca2+ dependent manner. It is a very well-studied function required during transmitter uptake and release. The rate of release can be plotted against time and then a time when release is minimum can be used for the intracellular glutamate assay. Otherwise, total glutamate in medium + cells can be measured.
What is the detection sensitivity of the assay? My media contains 0.05mM glutamate and I am looking for changes probably in the nM range. Is there another kit beter suited for this purpose?
The highest limit of detection is 1000uM or 1 mM and the lowest limit of detection is 40uM. So 50uM glutamate in the medium should be detectable by this kit.
If I run low on developer mix, s it possible to cut the amount of developer and enzyme mix in half and volume up the buffer to still have 100ul per well?
If you wish to use only half the Developer volume, it would require scaling down everything into half the volume to maintain relative concentrations.With the decrease in volume recommended here, absorbance values produced should not be directly compared to previous wells that contained full volume since absorbance depends on the path length of light through the sample.
Can cells lysed in RIPA buffer be used with this kit?
RIPA buffer typically contains SDS which might affect the function of the enzymes in the kit. Hence we do not recommend using RIPA bufefr samples. The assay buffer in the kit also provides optimum conditions for the enzymes to work at their best.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
How are samples normalized against protein concentration?
A protein quantitation assay can be used with the supernatants from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can individual components of this kit be purchased separately?
Yes, any of the kit's components can be purchased separately without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Wendy Xin, Oligodendrocytes Support Neuronal Glutamatergic Transmission via Expression of Glutamine Synthetase. Cell Rep., May 2019; 31116973.
Liu, F. et al., (2017) MicroRNA-200c exacerbates the ischemia/reperfusion injury of heart through targeting the glutaminase (GLS)-mediated glutamine metabolism, European Review for Medical and Pharmacological Sciences, 2017, 21: 3282-3289
Mus F et al., (2017) Diazotrophy overcomes the deleterious growth phenotype of glnE deletion in Azotobacter vinelandii, Appl. Environ. Microbiol., 2017, April 21: epub
Jin et al., Rett Syndrome Microglia: A Mechanism for Mitochondrial Dysfunction and Neurotoxicity. J. Neurosci., Feb 2015; 35: 2516 - 2529.
Gu et al., Erythropoietin Exerts a Neuroprotective Function Against Glutamate Neurotoxicity in Experimental Diabetic Retina. Invest. Ophthalmol. Vis. Sci., Dec 2014; 55: 8208 - 8222.
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