PrincipleThe kit is used to detect Glutamine in cell culture medium by measuring the absorbance of WST formazan produced according to quantities of glutamine. The kit includes a “Glutamine Standard” which can be used to quantify the concentration of lactate found within samples by creating standard curve.
Simple Assay Procedure
*The glutamine concentration can be calculated by using the following equation:glutamine (mmol/l) = (Glutaminase solution (+))－(Glutaminase solution (-))
Standard CurveStandard curve can be prepared by using Glutamine Standard included in the kit. The concentration of Glutamine can be measured. The concentration of glutamine can be evaluated by diluting the samples if the concentration is over 0.5 mmol/l.
Example of Measurements of Glutamine/Glutamate levelA549 cells were seeded on 6-well plates. Glutamine concentrations in the culture supernatant over incubation time were measured using the Glutamine Assay Kit-WST, and changes in glutamate concentration were measured using the Glutamate Assay Kit-WST. As a result, upregulated glutaminolysis led to a time-dependent decrease in glutamine concentrations and a time-dependent increase in glutamate concentrations in culture medium.
The number of samples that can be recorded when the standard curve and sample is measured in triplicates.100 testsSample 12 samples*(measured in triplicate – See picture below)
A plate layout for Glutamate standard solution and sample
Prepare a test sample to be assayed. Please note that a minimum of 240 μl of sample (e.g., supernatant, lysate) is needed for each test sampleThe glutamine concentration can be calculated by using the following equation:glutamine (mmol/l) = (Glutaminase solution (+))－(Glutaminase solution (-))
You can’t store the Working solution. Please prepare the Working solution prior to use. Please protect from light because the Working solution is light-sensitive.
No, the kit is used for the quantification of L-Glutamine levels.
You can’t accurately measure Glutamine levels if your samples contain a reducing agent. WST dye interferes with the reducing agent and can cause errors in observations. If you add a testing material that has reducing properties, please prepare a few wells that only contains the medium (no cells) and testing material for background control.