Glutamate is an important neurotransmitter that contributes to the biosynthesis of protein and glutathione. An excess of glutamate is considered to be a cause of neurodegenerative disorders, including Alzheimer's disease. A recent study showed that an iron-dependent cell death called “ferroptosis” is induced by inhibition of cystine/glutamate antiporter (xCT) responsible for cystine uptake and glutamate release. As such, cancer research which targets xCT has also been conducted in recent years.
Our Glutamate Assay Kit-WST is designed to quantify glutamate as a metabolite. This kit allows you to quantify glutamate present in culture medium or intracellular glutamate by WST reduction reaction. The lowest concentration of glutamate which can be quantified is 5 μmol/l. This kit can be used with 96-well microplates, which makes it possible to analyze multiple specimens.
The kit is used to detect Glutamate in cell culture medium by measuring the absorbance of WST formazan produced according to quantities of glutamate. The kit includes a “Glutamate Standard” which can be used to quantify the concentration of lactate found within samples by creating standard curve.
Simple Assay Procedure
Procedures are so easy that you simply incubate the plate after the addition of culture supernatant or tissue/cell lysates prior to adding a reagent.
Standard curve can be prepared by using Glutamate Standard included in the kit. The concentration of Glutamate can be measured. The concentration of lactate can be evaluated by diluting the samples if the concentration is over 0.5 mmol/l.
Example of Measurements of Glutamine/Glutamate level
Example of Measurements of Glutamine/Glutamate level in Ferroptosis research
It is known that an iron-dependent cell death called “ferroptosis” is induced when xCT is inhibited by elastin treatment. The amounts of glutamate release and intracellular glutathione were measured by using the elastin-treated A549 cells. As a result, the amount of glutamate release was decreased in the elastin-treated A549 cells, and the amount of intracellular glutathione was decreased by inhibiting cystine uptake.
Q. How many samples can I measure?
Ａ.The number of samples that can be recorded when the standard curve and sample is measured in triplicates.
(measured in triplicate)
(See picture below)
A plate layout for Glutamate standard solution and sample
Q. Can I quantify D-Glutamate levels?
A. No, the kit is used for the quantification of L-Glutamate levels.
Q. Can I measure samples that contain a reducing agent?
A. You can’t measure an accurate Glutamine level because WST dye interferes with the reducing agent. If you add test material which possesses reducing properties, please prepare a few wells that only contain the medium without cells and only test material for background control.
Q. Our samples did not change in color, are there any reasons for this?
Ａ. Glutamine level contained in the sample may be lower than the detection limit (5 µmol/l) that can be measured using this kit. If the lactate level is below 5 µmol/l.
If you dilute the sample, lactate level contained in the diluted sample may be lower than 5 µmol/l.
Please lower the dilution ratio and adjust the concentration of the sample over the detection limit that was measured using this kit.