Glutamate Assay Kit-WST
Glutamate Assay
- Easy extraction of cell Lysate
- Used to determine intracellular levels of Glutamate
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Product codeG269 Glutamate Assay Kit-WST
| Unit size | Price | Item Code |
|---|---|---|
| 100 tests | ＄529.00 | G269-10 |
| 100 tests | ・Dye Mixture ・Glutamate Standard ・Enzyme ・Assay Buffer ・Reconstitution Buffer ・lysis Buffer ・Filtration Tube |
×1 300 ×1 ×1 7.5 ml×1 750 ×l×1 25 ml×1 ×24 |
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Description
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.
Manual
Technical info
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
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
A549 cells were seeded in 6-well plates and the concentration of glutamine and glutamate in the culture supernatant were measured over time using the Glutamine Assay Kit-WST and Glutamate Assay Kit-WST, respectively.
The results showed that the concentration of glutamine in the culture medium decreased with time, while the concentration of glutamate increased with time.
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.
Sulfasalazine Alters Intracellular Metabolites and Increases ROS in A549 Cells
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After addition of sulfasalazine (SSZ), a known inhibitor of cystine/glutamate transporter (xCT), to A549 cells, we observed changes in intracellular glutathione (GSH), ATP, α-ketoglutarate (α-KG), ROS and glutamate release. The results indicated that the addition of SSZ
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Q & A
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Q
How many samples can I measure?
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A
The number of samples that can be recorded when the standard curve and sample is measured in triplicates.
100 Tests
Sample 24 samples*
(measured in triplicate – See picture below)
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Q
Can I quantify D-Glutamate levels?
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A
No, the kit is used for the quantification of L-Glutamate levels.
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Q
Can I measure samples that contain a reducing agent?
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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.
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Q
Our samples did not change in color, are there any reasons for this?
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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.
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Q
How stable is Working solution?
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A
Working solution cannot be stored. Prepare as needed. Because the working solution is unstable under light, it should be protected from light after preparation.
*Working solution is stable for 4 hours at room temperature if protected from light after preparation. If exposed to light, the color of the solution will turn brownish.
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Q
Can the samples be stored?
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A
We have confirmed that cell culture supernatant samples can be stored for one month at -20°C.
For cell lysate samples, we have confirmed that they can be stored for one month at -20°C. Before storing, however, please remove protein by using the Filtration Tube provided in the kit.
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Q
Is it possible to measure with absorption filters other than 450 nm?
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A
Other than 450 nm, a 490 nm filter can be used.
However, the absorbance value will be lower than when measured at 450 nm.
Handling and storage condition
| 0-5°C, Protect from light, Protect from moisture |
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